sero-prevalence and risk factors of infectious bovine
TRANSCRIPT
SERO-PREVALENCE AND RISK FACTORS OF INFECTIOUS BOVINE
RHINOTRACHEITIS IN THE SMALLHOLDER DAIRY FARMS OF NAARI SUB-
LOCATION OF MERU COUNTY, KENYA
A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF REQUIREMENTS FOR THE
AWARD OF THE MASTER OF VETERINARY MEDICINE (M. VET. MED), OF
UNIVERSITY OF NAIROBI
DR. SEREM ESSAU KIPYEGO
J56/8276/2017, (BVM)
DEPARTMENT OF CLINICAL STUDIES
FACULTY OF VETERINARY MEDICINE
COLLEGE OF AGRICULTURE AND VETERINARY SCIENCES
UNIVERSITY OF NAIROBI
NOVEMBER, 2019
ii
DECLARATION
This thesis is my original work and has not been presented for a degree in any other University.
Signature………………………………… Date…………………………………
Dr. Essau Kipyego Serem, (BVM, University of Nairobi)
This thesis has been submitted for examination with our approval as university supervisors
Signature………………………………. Date…………………………………
Prof. George K. Gitau (BVM, MSc & PhD)
Professor, Department of Clinical Studies, Faculty of Veterinary Medicine, University of Nairobi
Signature………………………………. Date…………………………………
Dr. Tequiero Okumu Abuom (BVM, MSc & PhD)
Lecturer, Department of Clinical Studies, Faculty of Veterinary Medicine, University of Nairobi
Signature………………………………. Date………………………………...
Prof. Daniel W. Gakuya (BVM, MSc & PhD)
Associate Professor, Department of Clinical Studies, Faculty of Veterinary Medicine University
of Nairobi
iii
DEDICATION
I dedicate my thesis work to my family and academic friends. A special gratitude to my loving
father, Joseph Kipserem Seroney and mother, Susana Jeruto for their constant prayer for my
wellbeing while undertaking my fieldwork and words of encouragement. I also dedicate my
dissertation to Non-Governmental Organization, Farmers Helping Farmers, the University of
Prince Edward Island and the University of Nairobi that has an existing developmental
partnership with the Naari Dairy Cooperative Society, for providing me with a strong foundation
for my study and also entry point to the community.
iv
ACKNOWLEDGEMENTS
I would like to acknowledge all the members of the Department of Clinical Studies, Faculty of
Veterinary Medicine, for the constant support they gave me through their expertise and precious
time. I would also acknowledge the developmental partnership between Farmers Helping
Farmers, the University of Prince Edward Island and the University of Nairobi for their full
support towards the research project. A special thanks to Prof. George Karuoya Gitau, lead
supervisor for linking me with Farmers Helping Farmers Organization, his countless hours of
reflecting, reading, encouraging, and most of all patience throughout the entire process. Thank
you Dr. Wycliff Ngetich, Dr. Willy Mwangi, Dr. Daniel Muasya, Dr. Nthenya Mbindyo, Dr.
Ambrose Kipyegon, Dr. Peter Kimeli, Dr. Gilbert Kirui, Dr. J. M. A. Kitaa, Prof Erastus Mutiga
and Prof John Vanleewan for constant guidance and developing my skills. I would like to
acknowledge the chairman of the Department of Clinical Studies, for allowing me to conduct my
research work and provision of any assistance requested. Special thanks to all members of staff
Biochemistry laboratory for their support and participation in this study. Finally, special thanks
to Dr. Tequiero Okumu Abuom and Prof Daniel Waweru Gakuya, my other supervisors, who
assisted me with this project. Their willingness to provide feedback made the completion of this
research work successful and a good experience.
v
TABLE OF CONTENTS
DECLARATION .......................................................................................................................... ii
DEDICATION ............................................................................................................................. iii
ACKNOWLEDGEMENTS ........................................................................................................ iv
TABLE OF CONTENTS ..............................................................................................................v
LIST OF TABLES ...................................................................................................................... vii
LIST OF FIGURES ................................................................................................................... viii
LIST OF APPENDICES ............................................................................................................. ix
LIST OF ABBREVIATIONS .......................................................................................................x
ABSTRACT .................................................................................................................................. xi
CHAPTER ONE: INTRODUCTION ..........................................................................................1
1.1. General objective .................................................................................................................. 2
1.2. Specific objectives ................................................................................................................ 2
1.3. Problem statement ................................................................................................................ 3
1.4. Justification .......................................................................................................................... 3
CHAPTER TWO: LITERATURE REVIEW .............................................................................5
2.1: Etiology of Infectious Bovine Rhinotracheitis ..................................................................... 5
2.2: Epidemiology of Infectious Bovine Rhinotracheitis ............................................................ 5
2.3: Immune Mechanism and Latency ........................................................................................ 7
2.4: Economic Importance .......................................................................................................... 8
2.5: Clinical Presentation of Infectious Bovine Rhinotracheitis ................................................. 9
2.6: Diagnosis of Infectious Bovine Rhinotracheitis ................................................................ 10
2.7: Differential diagnosis ......................................................................................................... 12
vi
2.8: Treatment of Infectious Bovine Rhinotracheitis ................................................................ 13
2.9: Prevention and Control of Infectious Bovine Rhinotracheitis ........................................... 13
2.9.1: Natural Exposure and Vaccination .............................................................................. 13
2.9.2: Biosecurity ................................................................................................................... 14
2.9.3: Closed herd .................................................................................................................. 15
CHAPTER THREE: METHODS AND MATERIALS ...........................................................16
3.1: Description of the study area.............................................................................................. 16
3.2: Selection of study area and farms ...................................................................................... 18
3.3: Data and sample collection ................................................................................................ 18
3.4: Laboratory analysis of samples using ELISA kits ............................................................. 19
3.5: Data Entry and Statistical Analysis .................................................................................... 21
CHAPTER FOUR: RESULTS ...................................................................................................23
4.1: Farm and animal demographics ......................................................................................... 23
4.1.1: Farm Demographics .................................................................................................... 23
4.1.2: Cow variables .............................................................................................................. 27
4.2: Management practices at the farms .................................................................................... 30
4.3: Sero-prevalence of Infectious Bovine Rhinotracheitis ...................................................... 32
4.4: Factors associated with Infectious Bovine Rhinotracheitis sero-prevalence in univariable
analysis ...................................................................................................................................... 34
CHAPTER FIVE: DISCUSSION ..............................................................................................38
CHAPTER SIX: CONCLUSIONS AND RECOMMENDATION .........................................43
6.1 Conclusions ......................................................................................................................... 43
6.2: Recommendations .............................................................................................................. 43
REFERENCES ............................................................................................................................44
APPENDICES ..............................................................................................................................55
vii
LIST OF TABLES
Table 4-1: Description of categorical variables for animal level factors for 403 cows on 149
smallholder dairy farms in Meru County, Kenya in 2018 ......................................... 28
Table 4-2: Description of continuous variables for animal and farm level factors (n = 403) on 149
smallholder dairy farms in Naari sub-location, Meru County, Kenya in 2018 .......... 29
Table 4-3: Description of categorical variables for farm level factors, 403 cows in 149
smallholder dairy farms in Naari sub-location, Meru County, Kenya in 2018 .......... 31
Table 4-4: Description of categorical variables for animal and farm level factors for 403 cows on
149 smallholder dairy farms in Meru County, Kenya in 2018 .................................. 33
Table 4-5: Univariable logistic mixed models of the outcome variable IBR type 1 antibody
seropositivity, while accounting for clustering of 403 cows among 149 smallholder
dairy farms in Meru County, Kenya in 2018, for variables of interest with P – Value
≤ 0.3 ........................................................................................................................... 35
Table 4-6: Final multivariable logistic mixed model for variables associated with IBR antibody
seropositivity for 403 cows on 149 smallholder dairy farms in Meru County, Kenya
in 2018........................................................................................................................ 37
viii
LIST OF FIGURES
Figure 3-1 A map showing Naari Sub-location in Meru County, Kenya. ................................... 17
Figure 4-1 The distribution of the principal dairy farmers Naari sub-location, Meru County,
Kenya in 2018 ............................................................................................................ 24
Figure 4-2 The distribution of the principal dairy farmers by age in Naari sub-location, Meru
County, Kenya in 2018 .............................................................................................. 25
Figure 4-3 The distribution of the dairy farmers by training on dairy production in Naari sub-
location, Meru County, Kenya in 2018 ...................................................................... 26
ix
LIST OF APPENDICES
Appendix 1: Descriptive statistics of continuous variables and proportion among the principal
farmers in 149 smallholder dairy farms in Naari Sub-location, Meru County, Kenya in
2018 ................................................................................................................................... 55
Appendix 2: Questionnaire for Management and Feeding Practices on Naari Smallholder Dairy
Farms ................................................................................................................................. 56
x
LIST OF ABBREVIATIONS
IBR Infectious Bovine Rhinotracheitis
BoHV Bovine Herpesvirus Virus
BVDV Bovine Viral Diarrhoea Virus
OIE World Organization of Animal Health
SAS Statistical Analysis Software
NDC Naari Dairy Cooperative Society
gB Glycoprotein B
ELISA Enzyme linked immuno-Sorbent Assay
PCR Polymerase Chain Reaction
VNT Virus Neutralization Test
CpHV – 1 Caprine herpesvirus – 1
CvHV – 1 Cervine herpesvirus – 1
CvHV – 2 Cervine herpesvirus - 2
DNA Deoxyribonucleic Acid
DIVA Differentiation of Infected from Vaccinated Animals
OD Optical Density
TMB Tetramethylbenzidine substrate
LM Light Microscope
IHC Immunohistochemistry
IgM M Immunoglobulin
IgG Gamma Immunoglobulin
BPIV – 3 Bovine Para-Influenza Virus Type 3
xi
ABSTRACT
The aim of the study was to analyze the sero-prevalence and risk factors of the Infectious Bovine
Rhinotracheitis disease among organized small holder dairy farms in the Naari area of Meru
County, Kenya.
A cross-sectional study was conducted in the Naari area of Meru County, Kenya between June-
July 2018 and March-April 2018. The 149 farmers were randomly selected from members of the
Naari Dairy Farmers Cooperative Society who were actively delivering milk to the society at the
time of the study. Serum samples were obtained from 403 female dairy cattle. Farm level
management and animal factors were collected through direct interviews with the owner or
someone who was knowledgeable about the animals. All serum samples were processed with an
indirect enzyme-linked immunosorbent assay (gB ELISA) to determine the presence of
antibodies to BHV-1.
The results revealed that the sero-prevalence of IBR among the smallholder dairy cows’
population in part of the large Meru County was 17.37% (95% CI: 13.80% to 21.43%). Among
the categorical predictor, the sero-prevalence of the breeds of the dairy cows were Ayrshire
(20.0%), Friesian (16.3%), Guernsey (17.9%) and Zebu (16.1%). The proportions positive for
BoHV -1 among parity of the dairy cows were Heifer (parity 0) (15.0%), Primi-para (parity 1)
(12.8%) and Multi-para (parity 2 – 8) (19.8%). The cow-heifer category was cow (18.7%) and
heifer (12.2%). The feeding systems employed in the production of dairy cows were zero –
grazing (17.5%), semi zero – grazing (18.9%) and grazing (12.1%). The study found out that the
IBR infection was positively associated with the following factors; age of the dairy cattle (OR =
1.112, 95% CI = 1.012 – 1.222, P = 0.027), cows that were borrowed into farms (OR = 4.893,
95% CI, = 1.328 – 16.03,1 P = 0.017), rearing goats in the farms (OR, 1.438, 95% CI, 1.109 –
xii
1.863, P = 0.006), cows that were given out of the farms (OR, = 2.486 95% CI, = 0.697 – 8.859 p
= 0.014), Showed BVD signs OR = 1.243 95% CI, = 0.635 – 2.430, p = 0.526) and cows that had
antibodies against Bovine Viral Diarrhea Virus (OR, = 2.262 95% CI, = 1.129 – 4.533, P =
0.021). The final multivariate analysis of individual level factors that were associated with those
that tested positive to BoHV – 1 antibodies included; rearing goats on the farm (OR = 4.636,
95% CI = 2.053 – 10.467, P = 0.001), age of the dairy cattle, (OR = 1.113, 95%CI = 1.017 –
1.217, P = 0.020) and age of the female principal farmers (OR = 0.174, 95% CI = 0.082 – 0370,
P = 0.001).
The study concluded that BoHV – 1 is naturally circulating among cattle population in Meru
County, Kenya. There was a positive association between age of the dairy cattle, age of the
principal female farmers and rearing of goats in the farm together with cattle, and BoHV – 1
sero-prevalence observed in the study. Thus, cattle population may be protected via punctual
vaccination while considering the differentiation of infected from vaccinated cattle. Furthermore,
there is need for a study to be carried out to identify long term effects of BoHV – 1 and access
the potential ability of the viral cross infection with other four related Alpha-herpesviruses with
BoHV – 1 among the cattle population.
Keywords: Infectious Bovine Rhinotracheitis (IBR), Bovine Herpesvirus type 1 (BoHV – 1)
Cattle, Sero-Prevalence and risk factors.
1
CHAPTER ONE: INTRODUCTION
Infectious Bovine Rhinotracheitis disease is of significant economic importance worldwide
caused by Bovine Herpes Virus – 1 (BoHV – 1) and it affects both domestic and wild ruminants
(Bowland et al., 2000; Muylkens et al., 2007). Bovine Herpes Virus – 1 is a virus of genus;
Varicellovirus, subfamily; Alphaherpesvirinae and family; Herpesviridae is a highly contagious
and infectious virus (King et al., 2012; Biswas et al., 2013; Newcomer and Givens, 2016).
Various subtypes of the virus cause different syndromes in cattle. Infectious bovine
rhinotracheitis in bovine is caused by BoHV – 1.1, the respiratory subtypes. Strain BoHV – 1.2a
and BoHV – 1.2b are the genital subtype while BoHV 1.3 is the encephalitic subtype (Muylkens
et al., 2007).
Infectious bovine rhinotracheitis virus (IBRV) has potential for cross infection with other
ruminant Alpha-herpes viruses; Cervine herpesvirus – 1, Cervine herpesvirus – 2, bovine
herpesvirus – 1 and caprine herpesvirus – 1 (Yesilbag et al., 2003). Bovine Herpes Virus – 1 is
also closely related with elk herpesvirus and buffalo herpesvirus (Keuser et al., 2004). The virus
affects both domestic and wild ruminants; cattle, white-tailed deer, mule deer, water buffalo,
African wildebeest, roe deer, red deer, woodland caribou and reindeer. Goats are naturally
infected while pronghorn antelope and African buffalo are reservoirs of the disease. (Biswas et
al., 2013).
Infected cattle are the source of infection to the susceptible herds. These cattle shed the virus
through body secretions and excretions via nasal discharges or droplets, semen, genital
secretions, fetal fluid and tissues (Takiuchi et al., 2005; Constable et al., 2017). The virus is
transmitted through aerosol infection in the respiratory form and this is dependent on
environmental factors such as humidity and temperature. Direct contact with contaminated
2
semen, mucosal discharges, fetal tissues and fluid and genital discharges can also lead to
transmission (Mars et al., 1999; Mars et al., 2000; Kahrs, 2001).
Cattle of all breeds and ages are equally susceptible and the disease is common among cattle
above 6 months of age due to increased chances of exposure to the BoHV – 1 (Majumder et al.,
2015; Seyfi-Abad-Shapouri, et al., 2016; Constable et al., 2017). The disease has no seasonal
variability, though in temperate countries, during the months of fall and winter, the occurrence of
the disease is high due to feedlot cattle assembling (Majumder et al., 2015). Unvaccinated
breeding cattle or feedlot cattle are susceptible to epidemics of abortion and respiratory diseases.
The systemic type of the disease is common with newly born calves with inadequate colostrum
antibodies or failure of passive immunity (Constable et al., 2017). The managerial and
environmental risk factors contributing to the spread of BoHV – 1 include; purchasing of
infected cattle, participation in agricultural shows, increased herd size and production system.
Uncontrolled movement of visitors and cattle within the farm, and unreliable records of
vaccination dates also contributes to the spread of the disease (Boelaert et al., 2005; González-
Garcia et al., 2009).
1.1. General objective
The overall objective of this study was to investigate sero-prevalence and risk factors of
Infectious Bovine Rhinotracheitis among smallholder dairy cattle in the Naari area of Meru
County, Kenya
1.2. Specific objectives
This study was designed with the following specific objectives;
1. To determine the sero-prevalence of Infectious Bovine Rhinotracheitis virus in the
Naari area of Meru County, Kenya.
3
2. To assess the risk factors associated with Infectious Bovine Rhinotracheitis virus
infections in cattle in the Naari area of Meru County, Kenya.
1.3 Problem statement
Infectious Bovine Rhinotracheitis disease is of significant economic importance in the dairy
industry causing losses due to respiratory diseases, calf mortalities up-to 100% and reproductive
losses. Vaccination against the disease has probability to reduce the economic losses due to
clinical disease and not the prevalence of BoHV – 1 infection. However, most infected cattle
often show no clinical picture and it`s therefore difficult to correctly estimate the economic
impact of the clinically sick dairy cattle. The spread of the disease may vary within the country,
from one region or farm to farm due to stock densities, managerial differences, micro-climatic
changes and other factors. The risk factors of BoHV – 1 infections include; herd size,
introduction of new animals to the farm, season, production system, vaccination status and
animal age. Thus, information on epidemiology of Infectious Bovine Rhinotracheitis is vital in
development of prevention and control programmes. Although Infectious Bovine Rhinotracheitis
disease has been reported all over the world, there is scarcity of information about the disease in
Kenya and this was what this study was seeking to address.
1.4 Justification
Infectious Bovine Rhinotracheitis is a disease of economic importance in cattle. Cattle greatly
contribute to the economy and welfare of most Kenyan rural populations. The economic losses
are associated with treatment costs, inefficient feed conversion efficiency, reduced conception
rates, loss of newborn calves, low milk yield, abortion and loss of body condition (Constable et
al., 2017). This study was prompted by scarcity of information about the disease in Kenya. The
study aims at improving the knowledge on Infectious Bovine Rhinotracheitis in Eastern Africa
4
through estimation of sero-prevalence and the risk factors accompanying the disease. Some of
the economic gains from this studies will include reduced clinical cases, increased reproductive
and production turnover and forms part of epidemiological surveillance.
5
CHAPTER TWO: LITERATURE REVIEW
2.1: Etiology of Infectious Bovine Rhinotracheitis
Bovine herpesvirus type 1 is an alpha-herpesvirus of family; Herpesviridae, subfamily; Alpha-
herpesvirinae and genus Varicellovirus. Genetic analyses of clinical isolate found four distinct
types; BoHV – 1.1, BoHV – 1.2a, BoHV – 1.2b and BoHV – 1.3. BoHV – 1.3 being neuropathic
sero-type has about three genotypes; BoHV – 5a, BoHV – 5b and, BoHV – 5 non-a and non-b
(Mahony, 2010). The observed antigenic differences among isolated viruses account for the
diverse pathologic and epidemiologic pattern. However, vulvovaginitis/balanoposthitis or
rhinotracheitis depend on the infection route rather than the subtype of the virus.
The four cud-chewing hoofed mammals alpha-herpesviruses related with BoHV – 1, may have
potential for cross-infection with cattle include: bovine herpesvirus type 5 (BoHV – 5), cervine
herpesvirus type 1 (CvHV – 1), caprine herpesvirus type 1 (CpHV – 1) and/or cervine
herpesvirus -2 (CvHV – 2). Other alpha-herpesvirus which are closely related with BoHV – 1
include: elk herpesvirus and buffalo herpesvirus – 1. Bovine herpesvirus – 5 causes fatal type of
meningoencephalitis among the calves while CpHV – 1 causes generalized infection among
neonatal kids’ and enteritis, Cervine herpesvirus – 1 may cause ocular diseases among red deer
and is widespread in both free-living and farmed red deer and CvHV – 2 also has been isolated in
Finland among the reindeer (Constable et al., 2017). Bovine herpesvirus – 4 (BOHV – 1) has
been isolated from case of bovine mastitis (Constable et al., 2017.
2.2: Epidemiology of Infectious Bovine Rhinotracheitis
Infectious Bovine Rhinotracheitis disease has been recorded in various countries worldwide
including; Germany, Sweden, Finland, Italy, Norway, United States of America, Switzerland
Canada, Denmark and African countries (Muylkens et al., 2007). Norway, Sweden, Switzerland,
6
Finland, parts of Italy and Germany have managed to eradicate the virus (Ackermann & Engels,
2006; Raaperi et al., 2014). Denmark, Switzerland, Sweden, Finland and Austria are officially
free of Infectious Bovine Rhinotracheitis (Ackermann and Engels, 2006). However, the sero-
prevalence of Infectious Bovine Rhinotracheitis virus varies from continents, countries and
regions and this is caused by differences in climates, stocking densities and management among
other factors (Ackermann and Engels, 2006; Almeida et al., 2013). Sero-prevalence of BoHV – 1
varies from region to region with the rate of 64.4% being reported in North-Eastern Mexico
(Segura-Correa et al., 2016), 65.88% in Northern part of Tamil Nadu, India (Saravanajayam et
al., 2015), 36%–48% in South and Central America, 14%–86% in Africa (Straub, 1990; Ghirotti
et al., 1991; Mahmoud et al., 2009), 43%, England (Woodbine et al., 2009) and 36%, China
(Yan et al., 2008).
Previous studies of the disease in Western part of Kenya revealed sero-prevalence rate of IBR
was at 20.9% (Callaby et al., 2016). According to the study, they found out that, bovine para-
influenza virus type 3, IBR and BVD have an association (Fulton et al., 2000; Callaby et al.,
2016). Sero-prevalence of this disease among the small scale dairy cattle farms in coastal areas
of Kenya was 28.6%, with significant increase in seroconversion rates with increasing age
(Kenyanjui et al., 2007).
The reproductive type of the disease due to BoHV – 1 was reported in Germany as infectious
pustular balanoposthitis/vulvovaginitis (Segura-Correa et al., 2006; Abu Elzein et al., 2008;
Graham, 2013). The virulent type of the disease is associated with BoHV – 1.1 which causes
respiratory type of the disease. Morbidity and mortality of the disease is higher in feedlot cattle
due to congregation of the susceptible population and introduction of new susceptible cattle from
epizootic region (Graham, 2013). The case fatality and morbidity rates among the dairy cattle are
7
3% and 8% respectively. In feedlot cattle, morbidity rates of between 20 – 30% have been
recorded (Majumder et al., 2015). A case fatality rate of less than 1% has been recorded in
feedlot animals though it may reach 10% with secondary bacterial bronchopneumonia and
tracheitis, (Graham, 2013; Constable et al., 2017).
2.3: Immune Mechanism and Latency
Immune response to the virus consists of local and systemic cell-mediated and antibody
immunity. The initial immune response to the virus in cattle exposed experimentally result in
release of IgM or IgG antibodies. The second wave of immune response results from abortion
following intra-amniotic inoculation of the virus leading to increase in IgM antibodies. Intranasal
exposure of the virus does not produce IgM antibodies (Bahari et al., 2013; Ghaemmaghami et
al., 2013; Haji-Hajikolaei and Seyfi-Abad-Shapouri, 2006). Once the cattle are infected naturally
or vaccinated with modified live-viral vaccines, the humoral and cell-mediated immune systems
are activated (Winkler et al., 1999; Jones, 2009).
Following intranasal infection or vaccination, antibodies and interferon production appears from
the third day and persist up to ten days. Humoral immunity has been used to indicate previous
bovine herpes virus – 1 infection (Constable et al., 2017). Cattle with low antibody may have
immunity due to cell mediated immunity. Evaluation of the cell mediated immunity as per the
previous studies may be done using delayed hyper-sensitivity test (Constable et al., 2017).
However, BoHV – 1 may become latent after primary infection or vaccinated with attenuated
viral strains. The location of the viral latency in cattle body varies; localized on replication site,
sacral or trigeminal ganglion (Seyfi-Abad-Shapouri, et al., 2016; Constable et al., 2017). BoHV -
1 have been isolated in about 10% of clinically healthy cattle on the trigeminal ganglia at the
8
slaughter, 40% of the bovine had serum neutralizing antibodies to the BoHV – 1 (Winkler et al.,
2000; Constable et al., 2017).
Recrudescence, rise in neutralizing antibodies and viral shedding, occur following exposure of
cattle to stress factors; high doses of corticosteroids, parturition, transportation and high ambient
temperature (Radostits et al., 2000; OIE, 2008; Viu et al., 2014). Detection of the latency form of
the BoHV – 1 among cattle populations is vital in control and prevention practices, and
international trading activities. Thus, tests to assay specific antibodies in the sera samples should
have a higher sensitivity to detect low viral specific antibodies with emphasizes on international
standardization of the tests (Moore et al., 2000; OIE, 2008). In endemic herds, transmission of
the virus is non-continuous, however, its sufficient to obtain detectable quantities of antibodies.
Latent infection result in negative serological test, since no animal reinfection to stimulate
humoral immunity (Geraghty et al., 2012).
2.4: Economic Importance
Infectious Bovine Rhinotracheitis causes significant economic impact in both beef and dairy
cattle reproduction and production feedlots. The losses incurred as a result of infertilities are due
to infectious pustular balanoposthitis in male cattle and infectious pustular vulvovaginitis in
cows. Other losses may include; epidemic abortion, production loss, deaths due to respiratory
disease among all ages of the cattle, deaths among newborn calves due to highly fatal form of
systemic diseases and cost of disease management due to secondary bacterial infection of the
respiratory system occurs (Bowland & Shewen, 2000; Saravanajayam et al., 2015; Constable et
al., 2017).
9
2.5: Clinical Presentation of Infectious Bovine Rhinotracheitis
Clinically, IBR presents with fever (39.9 – 420C) and sudden drop in milk yield which eventually
cease completely for 24 – 48 hours. Other signs include; hyper-salivation, inflamed mucous
membranes with nasal discharge that is initially mucoid then later mucopurulent, short course of
cough, apathy and anorexia (Mulyken et al., 2007; Graham, 2013).
The respiratory disease affects oro-respiratory mucosa and the clinical picture includes; anorexia,
fever, excessive salivation, coughing and nasal discharge, inflamed nares, conjunctivitis and
lacrimal discharge. BoHV – 1.1 replicates on epithelial cells thus kills the respiratory mucosal
cells thus, causing damage and necrosis of epithelial tissues (Jones, 2009).
The virus may also affect CD4+ T lymphocyte cells, affecting antigen production mechanism
and CD8+ T lymphocyte cells recognition mechanism among the infected cells (Winkler et al.,
1999; Koppers-Lalic et al., 2001). Furthermore, the dampening host-mounted an interferon
response by employing the diverse modes of strategies (Henderson et al., 2005; Saira et al.,
2007; Saira et al., 2009; Da Silva et al., 2011 and Da Silva et al., 2012). The impaired non-
specific immunity of the host provides opportunity where respiratory tract commensal bacteria of
the family Pasteurellaceae may colonize the healthy lower respiratory tract system (Griffin et
al., 2010).
Abortion is a common sequela in reproductive disease. Abortion may occur several weeks after
vaccination or clinical illness of unimmunized pregnant cow using modified live-virus vaccines
of bovine tissue culture origin (Constable et al., 2017). moreover, abortion may occur up-to 90
days of pregnancy especially when the virus goes latent within placenta thereafter infects foetus
much later than usual. However, possibility of a vaccine to cause abortion even with safe
vaccines may occur if the natural infection preceded vaccination, this commonly occur between
10
6 and 8 months of pregnant cow (Constable et al., 2017). Retained after birth is often common
and may be followed by residual infertility. Short estrus, endometritis and poor conception rates
may occur if breed herd is inseminated with infected semen (Graham, 2013).
Infectious pustular vulvovaginitis manifests clinically with a mild vaginal discharge, elevated
tail, and frequent urination. The vulva may present with small papules, swollen, mucosal
ulceration and erosion on the surfaces. Ulcers on the mucosal surfaces may coalesce and slough
off resulting to brown necrotizing tissue surfaces. Recovery may occur in about 10 – 14 days
unless preceded by complications. On the other hand, balanoposthitis manifest clinically with
small pustules, erosion and ulcers on glans penis and preputial mucosal (OIE, 2010; Bosco
Cowley et al., 2011; Gould et al., 2013).
The ocular form of the disease presents with reddened and edematous conjunctiva, profuse
serous ocular discharge and diffuse edema. Calves of 6 months of age and below can develop
encephalitis accompanied with incoordination, excitement, high mortality rates, hyper-salivation,
bellowing, convulsion and blindness (Constable et al., 2017).
New born calves less than 10 days of age often come down with systemic type of the disease
which is severe and invariably lethal. Clinical findings are varied and include; fever, hyper-
salivation, sudden anorexia, rhinitis accompanied by either unilateral or bilateral conjunctivitis,
hyperemia of buccal mucous membranes, erosion of the soft palate covered with tenacious
mucus, acute pharyngitis covered with tenacious mucopurulent discharge, edematous larynx,
bronchopneumonia, diarrhea and dehydration (Constable et al., 2017).
2.6: Diagnosis of Infectious Bovine Rhinotracheitis
Experimental infection has revealed that the median period to shedding of the virus may be 2
days, median period towards peak viral shedding may be 4 days while the median period up to
11
shedding of the virus stops is 14 days (Grissett et al., 2015). Viral isolation from the nasal swabs
by employing tissue culture via combination of fourfold rise of antibodies either at acute or
convalescent phase of the sera are the desirable state for the positive diagnosis. Cotton or
polyester swabs samples are recommended for collecting nasal swabs compared to calcium
alginate swab sample because the latter is virucidal within two hours (Constable et al., 2017).
Viral detection using nasal swabs may involve the use of direct and indirect immunofluorescence
techniques, Enzyme Linked Immunosorbent Assay (ELISA), immune-peroxidase, or use of
electronic microscopic examination which show the herpesvirus-like viral particles.
However, sensitivity of the direct immunofluorescence techniques therefore, is comparable to
cell culture technique. In addition to, monoclonal antibodies detected by immunofluorescence
assay technique may discriminate the 4 ruminant alpha-herpesvirus related to BoHV – 1. The
ELISA technique used in detection of BoHV – 1 has a higher sensitivity compared to viral
neutralization test (Saravanajayam et al., 2015). Thus, combination of viral isolation and indirect
immunofluorescence test from both nasal and ocular swab samples from several cattle samples
will increase the chances of recovery rates (Roshtkhari et al., 2012 and Constable et al., 2017).
An alternative practical means of quick detection of the BoHV – 1 is the use of Polymerase
Chain Reaction (PCR) assay since it is as sensitive as viral isolation (Mahajan et al., 2013). PCR
assay are considered equivalent to the standard dot blot hybridization and/or virus isolation thus,
may be used also in viral detection in semen samples. The southern blot hybridization compared
to PCR assay has high sensitivity and may detect virus in semen samples before the virus
development of any detectable antibodies (Constable et al., 2017). The PCR assay may be able to
detect positive semen samples 5 times even in the virus isolated from egg yolk-extended semen.
12
Thus, PCR assay are considered the diagnostic test of choice especial in routine diagnosis of the
BHV – 1 in aborted fetuses (Mahajan et al., 2013).
Employing the restricted endonuclease enzyme analysis of the viral DNA may be possible in
differentiation of the field isolated virus from vaccine strains, thus, this is useful in investigation
of vaccines-induced epidemics of the disease (Constable et al., 2017).
The bulk–tank milk testing for BoHV – 1 antibody is important in control program, eradication
programs and monitoring programs since it provide a rapid inexpensive screening of the cattle.
However, correlation between within herd prevalence and bulk milk testing seropositive of the
cattle may be higher at about 0.86. When BoHV – 1 is detected in bulk milk test technique, thus
there could be a probability that more than one cattle in a herd may be infected and the infections
has spread (Constable et al., 2017).
The following samples are collected for confirmatory diagnosis of BoHV – 1; histology samples
include; formaldehyde fixed samples of neonate or abortion: liver, rumen, kidney, esophagus,
trachea, lung, pharynx and adrenal glands, respiratory form: pharynx, trachea, nasal turbinate and
lungs, encephalic form; half of the mid-sagittally sectioned brain for LM and IHC. Virology
samples includes; neonate/abortion: kidney, rumen, lungs and liver, respiratory type; nasal swab,
trachea and lung, encephalic type; half of the mid-sagittally brain section for fluorescent
antibodies test, ISO and PCR (OIE, 2010; Constable et al., 2017 and Barber et al., 2017).
2.7: Differential diagnosis
IBR is defined by anorexia, acute rhinotracheitis, excessive nasal discharges, coughing, bilateral
conjunctivitis, nasal lesions, fever and gradual recovery within few days. However, secondary
form of pneumonia and bacterial tracheitis may occur. Thus, IBR should be differentiated from
bovine viral diarrhea, malignant head catarrh, pneumonic pasteurellosis, calf diphtheria, viral
13
pneumonia and allergic rhinitis (Griffin et al., 2010 and Constable et al., 2017). It`s important to
differentiate systemic disease of IBR in newborn calves among the following diseases; toxemias,
septicemia and acute pneumonia (Constable et al., 2017).
2.8: Treatment of Infectious Bovine Rhinotracheitis
Infected animals are isolated, identified and closely monitored especially for evidences of
secondary pneumonia and bacterial disease which may be preceded by anorexia and toxemia.
Treatment of tracheitis may be difficult, however broad – spectrum antibacterial are indicated for
secondary pneumonia and bacterial tracheitis. The antibiotics should be administered daily and
several days (Constable et al., 2017).
2.9: Prevention and Control of Infectious Bovine Rhinotracheitis
IBR being viral disease, can set in unpredictably at any period, even in closed herds, then sudden
outbreaks of the disease are experienced. The recent strategies of controlling IBR includes;
biosecurity measures, natural exposure or vaccination may be effective in eradication of the virus
in the herd of cattle population in a region or a country (Constable et al., 2017).
2.9.1: Natural Exposure and Vaccination
The cattle which recovered from natural infection of BoHV – 1 are immune against further
infections. However, naturally infected cattle are risky since all cattle population will not become
infected and obtain immunity to further clinical disease (Constable et al., 2017). Storm of
abortion occur in unvaccinated herd, thus, vaccination is recommended in region with high
prevalence and unfeasible eradication due to extensive nature of the cattle population and their
movement from one point to another across various region (Graham, 2013). The vaccination
rationale is normally determined by the following factors; the virus being ubiquitous and
occurrence of the diseases is unpredictable, the economic impact due to respiratory disease,
14
neonatal disease and abortion may be higher, maternal antibodies in newborn calves start to
wane at between 4 – 6 months of age and vaccination may prevent abortion and protect against
respiratory form of the disease if given at least 10 days before an animal is infected with the
virus naturally (Constable et al., 2017). Thus, vaccination of cattle as a mode of control and
eradication program as employed by the European countries was based on marker vaccine
deleted in gE gene. The inactivated or live attenuated marker vaccine employed in serological
diagnostic technique of detecting gE-specific antibodies, allowed discrimination of naturally
infected animals from vaccinated cattle (Van Oirschot et al., 1997; Lehmann et al., 2002). This
capacity employed in Differentiation of the Infected from Vaccinated Animals (DIVA) is key in
world trade restriction (Mars et al., 2001; Mulyken et al., 2007).
DIVA is demonstrated effectively with punctual vaccination of cattle at interval of six months’
apart, however, this technique is associated with a few weaknesses. Thus, the sensitivity of the
tool depends on capacity of the diagnostic test to detect the BoHV – 1 gE specific antibodies.
However, the diagnostic test sensitivity is readily available at around 70% using gE specific
ELISA technique (Perrin et al., 1996; Kramps et al., 2004). Another disadvantage is that the
response of the immune level raised against BoHV – 1 gE antibodies is weak thus, the window
period for the test ability to detect gE specific antibodies may be delayed up to 6 weeks (Beer et
al., 2003; Mulyken et al., 2007).
2.9.2: Biosecurity
This is an important measure in a successful livestock production since it reduces risk and effects
of introduction of infectious agents. The factors of biosecurity include; placement and
management programs, immunization, decontamination, farm layout and pest control (Constable
et al., 2017). Herpesviruses are sensitive to a number of disinfectants; 10% lugol`s iodine, 1%
15
phenolic derivatives and 1% quaternary ammonium bases (Constable et al., 2017). Introduction
of new infectious agents into herd may be minimized or prevented by purchasing new cattle from
farms known to be free of diseases in question. The adoption of this principles may require the
awareness of any possibility of unknown infectious agents including testing the cattle for
infectious agents before entry into the herd. It also involves quarantine of newly introduced cattle
for several weeks from arrival time to avoid mixing with clean herd (Constable et al., 2017).
Veterinarian play important role in development of specific disease control and biosecurity
protocols as per the farm or regional requirements (Constable et al., 2017). They also facilitate
development of methods of purchase of replacement stock and handling livestock through
designing known protocols concentrating on specific and general aspects likes designing and
putting up isolation rates (Constable et al., 2017).
2.9.3: Closed herd
Closed type of farming system facilitates prevention of emerging or re-emergence of infectious
agents into dairy cattle farms. Closed dairy farming enterprise may minimize introduction of
BoHV – 1, thus, this may form the baseline mechanism of eradication of infectious agents in
dairy herd (Constable et al., 2017).
Movement of cattle from one point to another; cattle shows and sales, veterinary clinics,
community grazing pasture, auction markets, club events, bull leasing and cattle commingling
from adjacent herds, provide opportunities for transmission of important infectious agents
(Constable et al., 2017).
16
CHAPTER THREE: METHODS AND MATERIALS
3.1: Description of the study area
A cross-sectional study was conducted in 149 smallholder dairy farms, Naari Area, Meru
County, Kenya (Figure 3.1). It lies at latitudes: 0°6'0" N and 37°34'60" E. Meru County is
located in the former Eastern Province of Kenya, 37o 18’37”to 37
o 28’33” E and 00
o 07’23” to
00o 26’19” S, approximately 270 km North of Nairobi, the capital city of Kenya. Meru shares
border with Isiolo County to the North, Laikipia County to the West, Tharaka Nithi and Nyeri to
the South West. The climate in Meru is warm and temperate. The average annual temperature in
Meru highlands ranges from14oC to 17
oC in the highlands while in the lowlands it ranges from
22oC to 27
oC. Precipitation in high altitude areas averages 2200 mm while low altitude areas
averages 500 mm. The Naari sub-location is situated in highly agricultural potential region
within an altitude of approximately 2000 m above the sea level. The main agricultural practice
includes; lumbering, horticulture, crop production and dairy production (Makau et al., 2018).
17
Figure 3-1: A map showing Naari Sub-location (middle of county) in Meru County of Kenya.
18
3.2: Selection of study area and farms
The study area was purposively selected since this research formed part of a larger study
involving smallholder dairy farmers (Figure 3.1). A non-governmental organization, Farmers
Helping Farmers, the University of Prince Edward Island and the University of Nairobi had an
existing developmental partnership with the Naari Dairy Cooperative Society, which provided a
strong foundation for this study and the entry point to the community.
The sampling frame for the study constituted of 568 farmers who were active members of the
Naari Dairy Cooperative (NDC) and shipping milk to the cooperative. The dairy cattle sampled
in the study was calculated based on an estimated prevalence of IBR of 50%, a precision of 5%
and confidence level of 95%, giving a sample size of 385 (Dohoo et al., 2009).
Since the average number of cattle per farm had been established as 2 to 3, then 149 farms were
randomly selected from the 568 smallholder dairy farms from the registry of active members
between January and June 2018 using software-based random number generation. The 149 farms
randomly selected provided about 400 animals.
3.3: Data and sample collection
The selected farms were visited between March-April 2018 and June and July 2018, and a
questionnaire was administered to capture farm and animal level factors (Appendix 1). The data
19
collected included collection of information about milking cows at the farm, systematic scrutiny
of written records to obtain the age of the cattle, calving rates, history of respiratory and
reproductive diseases, peri-parturient condition, and mastitis cases. Other information collected
included; feed and mineral supplementation, vaccination status, cattle owner attendances to any
dairy husbandry training, herd size, awareness and monitoring of heat signs, cow age and source
of animals.
In addition, 5 ml of whole blood in plain tubes for sera preparation was collected via the tail vein
of each dairy cattle, using 5 ml syringe and 21 gauge, 1.5mm needle. The blood tubes were
placed under shade to allow clot separation and thereafter the serum was transferred to
Eppendorf® vials which were labelled carefully, frozen and transported in ice to Heamatology
and Biochemistry Laboratory, Department of Clinical Studies, Faculty of Veterinary Medicine,
University of Nairobi and stored at -200C until testing.
3.4: Laboratory analysis of samples using ELISA kits
The frozen sera and ELISA kit from IDEXX Switzerland AG (Liebefeld-Bern, Switzerland)
which was stored at 40C, BHV – 1 Antigen Coated Plate and all the reagents, were thawed
carefully to room temperature (18 – 260C). The testing procedure was done following the
protocol described by manufacturer. The IDEXX IBR gB X3 was an indirect enzyme
immunoassay which has been developed to detect presence of antibodies against IBR in
individual bovine plasma, milk and serum samples. Antibody responses induced by vaccines
which contains the glycoprotein B (gB) of BoHV - 1 are detected as well. A microtration format
has been configured through immobilized IBR virus antigens on the plate. This kit is reported
with a specificity of 95% and sensitivity of 100%, and is able to detect the majority of BoHV - 1
antibodies.
20
Bovine herpesvirus – 1 Antigen Coated Plates were obtained and sample position was recorded.
Fifty microlitres of reconstituted wash solution was added to each well. Then 50 µl of Negative
and Positive Control samples was dispensed into respective labelled duplicate wells, and 50 µl of
each sample was dispensed into each respective sample well. The content of the micro-wells was
mixed via gently tapping the plate. The wells were hermetically covered with microplate cover
and incubated at 370C for 2 hours in a humid chamber. The solution was removed and each well
was washed with approximately 300 µl of wash solution for 5 times. The plates were protected
from drying between plate washings and prior to the addition of the next reagent. The final
washing of the plates was tapped on the absorbent material to remove any residual wash fluid.
Then gB specific monoclonal antibodies Horseradish Peroxidases conjugate was dispensed into
each micro-well, and incubated at 18 – 260C for 1 hour. The plate was rinsed as described above
and 100 µl of TMB Substrate N. 12 was dispensed into each micro-well.
The plate was then incubated at 18 – 260C for 10 minutes, and 100 µl of Stop Solution N. 3 was
dispensed on each micro-well. The test samples on antigen-coated well were incubated, and
antibodies specific to IBR virus formed complex with immobilized viral antigen. Unbound
antigen materials in the well were washed away, a gB-specific monoclonal antibodies
Horseradish Peroxidase conjugate was added. Thereafter, the unbound conjugate was washed
away and a substrate solution was added. The enzyme acted on the substrate converting it into a
product that reacted with chromogen to produce a blue color. The stop solution was thereafter
added resulting to the formation of yellow color.
The results were read from microplate photometer, Mindray Microplate Reader (MR-96A),
Shenzhen Mindray Bio-Medical Electronics Company Limited, where optical density (OD) was
measured either at a single wavelength of 450 nm [A (450)] or dual wavelength of 450 nm and
21
650 nm [A (450/650)]. The blocking percentage was calculated by using the absorbance [A
(450)] or [A (450/650)] obtained with the test sample and the negative control containing no
specific antibodies.
Interpretation of the results was determined via sample blocking percentage in accordance’s with
manufacturers test instructions where; blocking % < 45 negatives, 45 ≤ blocking % < 55 suspect
and blocking % ≥ 55 positives. The suspects were considered as positive in order to obtain a
dichotomous outcome.
3.5: Data Entry and Statistical Analysis
Data collected through the questionnaires and the laboratory results were first entered into MS
Excel (Microsoft Inc., Sacramento, California, USA) and then imported to Stata 15 (StataCorp
LLC, College station, Texas, USA) for analyses. Initially, the data were checked for accuracy,
coded and analyzed using descriptive statistics. Proportions were determined for categorical
variables, breed, age category, parity category and history of abortion, and presented as a
percentage of the overall number, along with a 95% confidence interval where applicable.
Mixed-effect logistic regression analysis was performed accounting for clustering of cows
among herds, to determine associations between the categorical variables, breed, age category,
parity category, history of an abortion, BVDV antibodies positive, showed BVDV signs, type of
feeding system, rearing goats on the farm, rearing sheep on the farm, use of natural mating,
fence-line contact with other cattle, grazing on community pasture, borrowed cows from other
farms, lent cows to other farms, cattle bought into the farm, dichotomized age of the female
principal farmers, dichotomized age of the male principal farmers, and continuous variables, age
of the animal, dry cows, herd size and milking cows, and the dichotomized seropositivity
22
outcome (presence or absence of IBR antibody). In the first step, univariable multi-level mixed
models for all the predictor variables were fitted into separate logistic regression models,
employing the functional logit. In the second step, a multivariable mixed logistic regression
analysis was fitted for all the univariable associations with p≤0.30 in the first step. Correlations
between predictors variables were identified using paired-wise correlation, and where two or
more variables were highly correlated (correlation coefficient >0.5), statistical significance and
biological plausibility were used to identify which variable would be offered to the modeling
process. The final models were built using backward stepwise elimination, leaving those
variables which had a p-value ≤0.05.
23
CHAPTER FOUR: RESULTS
4.1: Farm and animal demographics
4.1.1: Farm Demographics
The farm demographics are summarized in Appendix 1. A total of 403 cattle and 149 farms were
involved in the study. The principal farmer was mostly made of men (56.4%) while women were
fewer (28.2%), and 15.4% of farms had male and female considered as jointly principal farmer
(Figure 4-1). Most of the principal farmers were married (83.9%), but a few of them were young
people who were single and establishing themselves as dairy farmers (5.3%). Among the
principal farmers, a majority of the men were below 45 years (51.0%) while women also had the
majority of them above the age of 45 years (50.3%) (Figure 4-2). The large majority of the
female principal farmers had completed high school and tertiary school level of education
(84.9%), while the proportion of male principal farmers having completed high school and
tertiary school was slightly low at 79.8%.
The mean household size recorded in this study was 3.71 ± 1.54 with a minimum of 1 person and
a maximum of 11. The mean total land holdings ownership among the respondents was 2.11 ±
2.04 acres. In addition, some of the farmers (0.51 ± 0.84 acres) also had an access to other pieces
of land through leasing, borrowing and government owned lands lease to them.
The distribution of the training on dairy production among the principal farmers included those
with the dairy production training 81.2% (121/149) and those with no training on dairy
production 18.8% (28/149) with the majority of the principal farmer having no training on dairy
farming (Figure 4.3).
24
Figure 4-1 The distribution of the principal dairy farmers Naari sub-location, Meru
County, Kenya in 2018
Female Male Male/Female
Frequency 42 84 23
0
10
20
30
40
50
60
70
80
90
Fre
qu
en
cy
The distribution of principal farmers
The frequency of principal farmer
Frequency
25
Figure 4-2 The distribution of principal dairy farmers by age, Naari area, Meru County,
Kenya in 2018
Male ≥ 45 years Male < 45 yearsFemale ≥ 45
years Female < 45
years
Frequency 73 76 75 74
71.5
72
72.5
73
73.5
74
74.5
75
75.5
76
76.5
Fre
qu
en
cy
The distribution of the principal farmer by age
Frequency of the age of the principal farmers
Frequency
26
Figure 4-3 The distribution of the dairy farmers by training on dairy production in Naari
area, Meru County, Kenya in 2018
Trained farmersFarmers with no
Training
Frequency 121 28
0
20
40
60
80
100
120
140Fr
eq
ue
ncy
The distribution of principal farmer by dairy production training
Frequency of the principal farmer with dairy training
Frequency
27
4.1.2: Cow variables
Among the 149 smallholder dairy farms in the Naari Area, Meru County, 403 dairy cows were
recruited for the study. The animal-level variables are summarized in Tables 4-1 and 4-2. The
distribution of the study animals among the breeds kept in the region included; Friesian Holstein
47.2% (190/403), Guernsey 27.8% (112/403), Ayrshire 17.4% (70/403) and Zebu 7.7% (31/403).
Friesian Holstein formed the majority of the dairy cattle reared in the region.
The mean herd size was 5 with a range of 1-16 animals. Majority of the dairy cows in the farms
had a mean age of 5 years with a range of between 1 and 17 years. The average number of
milking animals per herd was 2 with a range of between 0 and 7 cows. The lactating cows
comprised the majority 79.7% (321/403) in the farms compared to replacement heifers/female
calves 20.3% (82/403) (Table 4-2). The parity of the cow was classified between 0 to 8 and
included, nulliparous heifer/female calves (parity 0) at 19.9% (80/403), primi-parous (parity 1) at
21.3% (86/403) and the multiparous (parity 2 and 8) at 58.8% (237/403). The farms had an
average number of dry cows of 1 with a range of between 0-3 animals per herd. In addition, the
farms had an average milking cows of 2 with a range of between 0-7. It was also reported that
about 20.1% (81/403) of the animals were reported to have experienced an abortion.
28
Table 4-1: Description of categorical variables for animal level factors for 403 dairy cattle on
149 smallholder dairy farms Naari area, Meru County, Kenya in 2018
Variable Category Category Frequency Percent by Category
Breed Ayrshire 70 17.4
Friesian 190 47.2
Guernsey 112 27.8
Zebu 31 07.7
Age category Cow 321 79.7
Heifer 82 20.3
Parity category Parity 0 80 19.9
Parity1 86 21.3
Parity >1 237 58.8
History of an abortion No history of abortion 322 79.9
History of abortion 81 20.1
29
Table 4-2: Description of continuous variables for animal and farm level factors (n = 403)
on 149 smallholder dairy farms in Naari area, Meru County, Kenya in 2018
Variable Range Mean S d Variance OR 95% CIOR
Age of the animal 5 - 17 5.521 3.240 10.496 1.112 1.012 – 1.222
Dry cows 0 - 3 0.149 0.516 0.266 1.446 0.685 – 3.049
Herd size 1 - 16 5.754 2.989 8.937 1.446 0.830 – 1.463
Milking cows 0 - 7 1.531 1.493 2.230 1.072 0.931 – 1.235
S – Standard deviation, IBR – Infectious Bovine Rhinotracheitis, OR – Odd Ratio & 95% CIOR –
95% Confidents Intervals of OR
30
4.2: Management practices at the farms
A summary of managerial practices found among the 149 smallholder dairy farms recruited in
the study are summarized in Table 4.3. The distribution of various types of feeding systems
among the smallholder dairy farms included; zero grazing system 42.2% (63/149), semi-zero
grazing system 35.7% (53/149) and grazing 22.2% (33/149). The zero-grazing and semi-zero
grazing systems formed the majority of smallholder dairy farms. Majority of the farmers
interviewed practiced zero-grazing with a few of the farmers grazing their cows in community
pastures 22.2% (33/149). Movement of the animals across the region were captured as follows:
cows that were borrowed into the farm 6.7% (10/149), cows that were lend out of the farm 8.1%
(12/149) and cows that were introduced into the farm 8.7% (13/149). Artificial insemination was
readily available in the region at 57.7% (86/149) and was offered by government veterinary
officers, veterinary technicians and private practitioners. Majority of the smallholder dairy farms
however used artificial insemination at 57.7% (86/149) while a few employed the natural mating
method at 42.3% (63/149). This may be associated with the high prices of semen, perceived low
conception rates and repeat breeding service. Among the farmers interviewed, other than dairy
production, they also practiced other production systems which included sheep 38.3% (57/149)
and goat 15.4% (23/149).
31
Table 4-3: Description of categorical variables for farm level factors, 403 dairy cattle in 149
smallholder dairy farms in Naari area, Meru County, Kenya in 2018
Variable Category Category
Frequency
Percent by
Category
Type of feeding system Zero– grazing 63 42.2
Semi-zero-grazing 53 35.6
Grazing 33 22.2
Rearing goats on the farm Yes 23 15.4
No 126 84.6
Rearing sheep on the farm Yes 57 38.3
No 92 61.7
Use of natural mating Yes 63 42.3
No 86 57.7
Fence-line contact with other cattle Yes 120 80.5
No 29 19.5
Grazing on community pasture Yes 98 65.8
No 51 34.2
Borrowed from other farms Yes 10 06.7
No 139 93.3
Lent to other farms Yes 12 08.1
No 137 91.9
Cows bought into the farm Yes 13 08.7
No 136 91.3
BVDV ab +vea Positive 157 53.4
Negative 137 46.6
Showed BVD signsb Positive 254 63.2
Negative 148 36.8
a Bovine Viral Diarrhea Disease Virus Antibodies positive cows
b Bovine Viral Diarrhea Disease
32
4.3: Sero-prevalence of Infectious Bovine Rhinotracheitis
The overall sero-prevalence of BoHV – 1 was 17.37% (70/403; 95% CI: 13.80% to 21.43%).
The sero-prevalence of the antibodies to the IBR virus in association to cow variables are
summarized in Table 4-4. For the categorical variables, the sero-prevalence for BHV-1 infection
inbreeds of the dairy cows were; Ayrshire 20.0% (14/70), Guernsey 17.9% (20/112), Friesian
16.3% (31/190) and Zebu 16.1% (5/31).
The sero-prevalence for BoHV-1 by parity of the dairy cows were primi-paraous 19.8%
(47/237), heifer 15.0% (12/80) and multi-parous 12.8% (11/86). Majority of categories detected
with IBR were cows 18.7% (60/321) while a few were heifers 12.2% (10/82). Majority of the
sero-positive animals were mainly reared under the zero-grazing and semi zero-grazing type of
production systems at 18.1% (21/116) and the sero-prevalence for open grazing was 12.1%
(4/33). The summary of the sero-prevalence to IBR to the other categorical variables are shown
in Table 4.4. The highest recorded sero-prevalence to IBR antibodies were observed in the
following variables: farms rearing goats 39.1% (9/23), farms rearing sheep 19.3% (11/57), cattle
grazing on the community pastures 19.4% (19/98), new animals introduced in the farms 23.1%
(3/13) and cows with negative antibodies against Bovine Viral Diarrhea Virus 23.4% (32/137).
33
Table 4-4: Description of categorical variables for animal and farm level factors for 403 dairy
cattle on 149 smallholder dairy farms, Naari area, Meru County, Kenya in 2018
Variable Category Category
Frequency
(Percent)
IBR
Percent + by
Category
Animal level factors
Breed Ayrshire 70 14(20.0)
Friesian 190 31(16.3)
Guernsey 112 20(17.9)
Zebu 31 05(16.1)
Age category Cow 321 60(18.7)
Heifer 82 10(12.2)
Parity category Parity 0 80 12(15.0)
Parity1 86 11(12.8)
Parity >1 237 47(19.8)
History of an abortion No history of abortion 322 55(17.1)
History of abortion 81 15(18.5)
Farm level factors
Type of feeding system Zero– grazing 63 11(17.5)
Semi-zero-grazing 53 10(18.9)
Grazing 33 4(12.1)
Rearing goats on the farm Yes 23 09(39.1)
No 126 17(13.5)
Rearing sheep on the farm Yes 57 11(19.3)
No 92 15(16.3)
Use of natural mating Yes 63 12(19.0)
No 86 14(16.3)
Fence-line contact with other
cattle
Yes 120 23(19.2)
No 29 03(10.3)
Grazing on community pasture Yes 98 19(19.4)
No 51 07(13.7)
Borrowed from other farms Yes 10 05(50.0) No 139 21(15.1)
Lent to other farms Yes 12 04(33.3)
No 137 22(16.1)
Cows bought into the farm Yes 13 03(23.1)
No 136 23(16.9)
BVDV ab +vea Positive 157 20(12.7)
Negative 137 32(23.4)
Showed BVD signsb Positive 254 41(16.1)
Negative 148 29(19.6)
34
4.4: Factors associated with Infectious Bovine Rhinotracheitis sero-prevalence in
univariable analysis
The study found out that the IBR sero-prevalence was positively associated with the following
factors; age of the dairy cattle (OR = 1.112, 95% CI = 1.012 – 1.222, P = 0.027), cows that were
borrowed into farms (OR = 4.893, 95% CI, = 1.328 – 16.03,1 P = 0.017), age of the female
principal farmers (OR = 0.230, 95% CI = 0.108 – 0.498, P = 0.000), age of the male principal
farmers (OR = 0.394 95% CI = 0.181 – 0.898, P = 0.019), rearing goats in the farms (OR, 1.438,
95% CI, 1.109 – 1.863, P = 0.006), cows that were given out of the farms (OR, = 2.486 95% CI,
= 0.697 – 8.859 p = 0.014) Showed BVD signs OR = 1.243 95% CI, = 0.635 – 2.430, p = 0.526)
and cows that had antibodies against Bovine Viral Diarrhea Virus (OR, = 2.262 95% CI, = 1.129
– 4.533, P = 0.021) (Table 4–5).
35
Table 4-5: Univariable logistic mixed models of the outcome variable IBR type 1 antibody
seropositivity, while accounting for clustering of 403 dairy cattle among 149 smallholder
dairy farms, Naari area, Meru County, Kenya in 2018, for variables of interest with P –
Value ≤ 0.3
Variable and category Categories OR 95% CIOR P–Value
Parity category 0.206!
Parity 0
Parity 1 1.059 0.361 – 3.103
Parity 2-8 1.414 0.803 – 4.428
Age category heifer
Cows 2.181 0.913 – 5.211 0.079
Fence-line contact with other cattle No
Yes 2.850 0.924 – 8.790 0.068
Grazing on community pasture No
Yes 2.150 0.918 – 5.033 0.078
Borrowed from the farms No
Yes 4.893 1.328–16.031 0.017
Lent to other farms No
Yes 2.486 0.697 – 8.859 0.014
Rearing Goats on the farm No
Yes 1.438 1.109 – 1.863 0.006
Rearing sheep on the farm No
Yes 1.076 0.969 – 1.192 0.173
Use of natural mating No
Yes 1.706 0.769 – 3.786 0.150
BVDV ab +vea No
Yes 2.262 1.129 – 4.533 0.021
Showed BVD signsb No
Yes 1.243 0.635 – 2.430 0.526
Age of the female principal farmers 0.230 0.108 – 0.498 0.001
Age of the male principal farmers 0.394 0.181 – 0.898 0.019
Age of the dairy cattle 1.112 1.012 – 1.222 0.027
Dry cows 1.446 0.685 – 3.049 0.249 ! Overall P-values for variables with >2 categories,
a Bovine Viral Diarrhea Disease Virus antibody
positive cows, b
Bovine Viral Diarrhea Disease, 95% CIOR: 95% Confidence Interval of OR, OR: Odds
Ratio
36
4.5: Factors associated with Infectious Bovine Rhinotracheitis sero-prevalence in
multivariable analysis
The final multivariate analysis of individual level factors that were associated with sero-
positivity to BoHV – 1 antibodies included; rearing goats on the farm (OR = 4.636, 95% CI =
2.053 – 10.467, P = 0.001), age of the dairy cattle, (OR = 1.113, 95%CI = 1.017 – 1.217, P =
0.020) and age of the female principal farmers (OR = 0.174, 95% CI = 0.082 – 0370, P = 0.001).
The likelihood of a older dairy cattle to have BoHV – 1 antibody is 1.113 times compared to
young dairy cattle. Farms that are rearing goats are 4.636 times more likely to have BoHV – 1
antibodies compared to farms not rearing goats. In addition, as the age of the women principal
farmer’s increases, the likelihood of a cow to have BoHV – 1 antibody were lower by 0.174
times compared with the younger women principal farmers (Table 4-6).
37
Table 4-6: Final multivariable logistic mixed model for variables associated with IBR
antibody seropositivity for 403 dairy cattle on 149 smallholder dairy farms, Naari area,
Meru County, Kenya in 2018
Variable OR 95% CIOR P – Value
Rearing goats on the farm 4.636 2.053 – 10.467 0.001
Age of the dairy cattle 1.113 1.017 – 1.217 0.020
Age of the female principal farmers 0.174 0.082 - 0370 0.001
OR: Odd Ratio, 95% CIOR: 95% Confidence Interval of OR
38
CHAPTER FIVE: DISCUSSION
The findings of this study confirmed the existences of IBR infection through sero-prevalence of
BoHV-1 antibodies among the smallholder dairy cattle reared in Meru County. Past studies in
Kenya have reported varying sero-prevalence’s to IBR for example a 20.9% in the western part
of Kenya and 28.0% in the former Malindi District of the Coastal Region were recorded
(Kenyanjui et al., 2007; Callaby et al., 2016). However, the sero-prevalence in this study was
lower than those reported and this could have been due to the different livestock production
systems studied and the way the studies were designed.
The observed sero-prevalence to Infectious Bovine Rhinotracheitis Disease of 17.4% was within
the range of 16% - 54% that was observed by McDermott et al. (1997) estimated in former
Districts in 1991 – 1992 in Kenya. However, the sero-prevalence observed in parts of Meru
County was lower than that which was observed in traditionally managed herds in Zambia,
which ranged from 42% -76% (Ghirotti et al., 1991) and in Egypt ranged from 63%–86%
(Mahmoud et al., 2009).
It has been observed that higher prevalence’s recorded in larger herd sizes and intensively
farmed cattle could be associated with a high level of contact between individual animals within
a herd (Snowder et al., 2006). For extensively managed farms with median herd size of 5
animals, the risk of contact between a susceptible individual with infected or persistently infected
animal is lower (Callaby et al., 2016). The studies reported above involved animals of various
ages for example 51 weeks old (Callaby et al., 2016), age 3 months to adults (Ghirotti et al.,
1991), zebu adults (Kenyanjui et al., 2007), and this study involved adult dairy cows and heifers.
The age differences among the animals studied may have explained some of the variations in the
sero-prevalence. In addition, Ghirotti et al. (1991) and Kenyanjui et al. (2007) employed virus
39
neutralization tests (VNT) compared to the ELISA tests used in this study and the differences in
test specificity and sensitivity may also have contributed to observed differences in prevalence
(Graham et al., 1998). In the studies done by Saravanajayam et al. (2015) found out that ELISA
has a higher specificity and sensitivity compared to VNT, thus ELISA is the rapid, reliable and
technically superior test for detection of BoHV – 1 antibody. Callaby et al. (2016) and IDEAL
study employed indirect ELISA test thus may have contributed to observe close proximity
prevalence’s. This observed close proximity prevalence’s may be attributed by use of ELISA
test, therefore similar in test sensitivity and specificity.
In other studies, in Africa, the IBR estimated prevalence among the cattle population ranged
from 14% to 86%, (Straub, 1990; Ghirotti et al., 1991; Mahmoud et al., 2009), whereas this
study revealed prevalence of 17.37% which is within the range. The observed differences in
antibodies prevalence in different regions, countries and counties could be explained by factors
likes production systems, differences in herd sizes; small, median or large herds, type of
breeding methods, differences in disease-control measures and age of the cattle, these are
important since they indicate diseases permanence in an environment (Orjuela et al., 1991;
Mainar-Jaime et al., 2001). Moreover, the observed differences in the IDEAL study and across
the world could be attributed by the breeding managements and geographical positions of the
considered in various regions and countries (Ackermann and Engels, 2006).
Cross-reaction may be observed between virus and its related viruses, for instances, BVDV has
potential for cross-reaction with pestiviruses like Classical Swine Fever and Border Diseases
Virus of sheep. Infectious Bovine Rhinotracheitis has the potential for cross-reaction with four
herpesviruses from other animals including goats and buffalo while the Para-Influenza Virus
type 3 (PIV3) can cross react with human strains of virus (Handel et al., 2011; Callaby et al.,
40
2016). Thus, since most of such viruses have not been exhaustively tested in cattle in Meru
County, the majority of sero-positive cases were likely to be due to exposure to IBR virus.
The fact that mixed rearing of sheep, goats and cattle is a common livestock production practice
in the region, there could be possibility of cross-reaction therefore the need for a study to be
carried out in the region in order to obtain more information on viral cross-infection. The present
study revealed that rearing of goats in the farm was highly associated with BoHV-1 sero-
prevalence. This may be attributed by the potential ability of the viral cross-infection with related
Alpha-herpesvirus (Biswas et al., 2013). In previous studies done by Whetstone and Everman
(1988), it was demonstrated that BoHV-1 has potential to infect and cause disease in goats and
sheep.
Based on past epidemiological studies, this study showed that IBR and BVD are closely
associated since one disease predispose to the other disease (Callaby et al., 2016). This finding
agrees with those reported earlier (Martin and Bohac, 1986; Durham and Hassard, 1990; Fulton
et al., 2000; Callaby et al., 2016). However, BoHV-1 infection is determined by stressor factors
such as use of steroids, transportation, parturition and high ambient temperature and these may
induce carrier animals to start shedding the virus (Six et al., 2001). Thus the clinical
manifestation of the disease depends on primary factors that lower the immunity system of the
animals (Seyfi Abad Shapouri, et al., 2016). This is because of the latency nature of the BoHV –
1 after primary infection or exposure to the virus or vaccination with attenuated viral strains
(Radostits et al., 2000; Constable et al., 2017).
The study also showed that BVDV sero-prevalence was significantly associated with BoHV-1.
Other studies found out positive correlation between IBRV, BPIV3 and BVDV (Callaby et al.,
2016). Martin and Bohac, (1986); Durham and Hassard, (1990) and Fulton et al., (2000), found
41
out that there was an association between the three viruses; BoHV-1, BVDV and PIV3.
However, Callaby et al. (2016) further noted that by including the environmental confounders,
the model quantified relationship between sero-positivity of IBRV, BPIV3 and BVDV but had
little effects on the association observed among the three viruses.
The findings of this study revealed that a number of risk factors such as age of the cattle, cattle
that were borrowed into farms, rearing goats in the farms, cows that were given out of the farms
and cows that had antibodies against Bovine Viral Diarrhea were associated with sero-prevalence
of BoHV – 1 antibody. The latter agree with several serological studies carried out worldwide to
identify risk factors associated with BoHV – 1 sero-positivity (Mulyken et al., 2007). The
current study also agrees with other studies which reported a number of risk factors in their
studies such as large herd size, housing, intensive production systems and managerial practices
like cattle movement and hygiene, age, sex with male frequently positive than females, direct
cattle contact especially participation in animal shows, purchasing of cattle and large herd sizes
(Van Schaik, 2001; Van Schaik et al., 2002; Solis-Calderon et al., 2003; Vonk Noordegraaf et
al., 2004; Boelaert et al., 2005).
Age of the cattle is a frequent risk factor reported in IBRV seropositive cattle, Solis-Calderon et
al. (2003), Cabonero et al. (2011), Romero-Salas et al. (2013), Saravanajayam et al. (2015) and
Segura-Correa et al. (2016) findings showed that older animals had a higher prevalence
compared to young animals while Saravanajayam et al. (2015) further reported that cattle over 3
years of age may have a higher sero-positivity than those of less than 3 years of age. The
findings of this study revealed that age of the cow is a significant risk factor to sero-prevalence
of BoHV – 1 antibody. This similarity could be attributed by the physiology of the cattle since
the ability of aging animal’s immunity system to defend against infection wears off compared to
42
young animals. Callaby et al. (2016) reported sero-positivity of 20.9% among the East African
Shorthorn Zebu calves, thus maternal antibodies against IBR are transferred to calves which
provides initial protection before development of the innate immunity. These observed
differences may be associated to animal inclusion criteria in the studies, for instance Callaby et
al. (2016) in his studies selection was based on calves age between 3 to 7 days old, the dam must
have been in the farm for not less than one year, and calf was due to natural mating.
The present study showed that introduction of new animals into the herd was significantly
associated sero-positivity of BoHV-1 hence this was an important risk factor. This difference
may be explained by differences in geographical location, disease-control programs employed
and herd sizes. Other past studies by Solis-Calderon et al., (2003) and Segura-Correa et al.,
(2016) reported that introduction or non-introduction of animal into the herd was not
significantly associated with sero-prevalence of BoHV-1. Furthermore, when the age of the
female principal farmer was over 45 years, the odds of cattle on the farm to have BHV-1
antibodies were lower by 0.174 times compared with cattle reared by younger women principal
farmers under or equal to 45 years (Table 3). This could be associated to majority of the female
principal farmers attend dairy production training sessions on husbandry, production,
reproduction and disease control training, thus, they obtain an understand on general disease
management in herds.
This study was carried out in smallholder farms with approximate herd size of 3 animals, as a
result, the study did not had enough power to identify herd size as risk factor for seropositivity.
Other studies suggested that at herd level, herd size is the main important risk factor for BHV-1
infection (Snowder et al., 2006; Segura-Correa et al., 2016). Our herd sizes were small, making
it less likely for our herds to be aggregates of animals from other farms.
43
CHAPTER SIX: CONCLUSIONS AND RECOMMENDATION
6.1 Conclusions
1. In Meru County, BoHV-1 is naturally circulating among the cattle population.
2. There was a positive association between IBR and age of the dairy cattle and rearing of
goats in the farm together with cattle.
3. Older women principal farmers had good experience with disease control methods, thus
the risk of diseases spread is reduced.
6.2: Recommendations
1. A study should be carried out to establish if the clinical IBR occurs in Naari sub location
in order to formulate prevention and control measures such as vaccination.
2. A study is also needed to be carried out to determine if their long-term effects of BoHV-1
sero-prevalence in cattle production.
3. Further study is required to access the potential ability of viral cross-infection with other
four related Alpha- herpesviruses with BoHV -1 which include; Cervine herpesvirus – 1,
Cervine herpesvirus – 2, bovine herpesvirus – 1 and caprine herpesvirus – 1.
44
REFERENCES
Abu Elzein E. M. E., Housawi F. M. T., Al-Afaleq A. I., & Al-Musa J. (2008). Emergence
of Clinical Infectious Bovine Rhinotracheitis in Eastern Saudi Arabia. Journal of
Animal Husbandry and Veterinary Medicine in Tropical Countries, 2008 61 (1): 11-13.
Ackermann, M. and Engels, M., (2006). Pro and contra IBR-eradication. Journal of
Veterinary microbiology 113(3-4): 293-302.
Almeida, L.L., Miranda, I.C.S., Hein, H.E., Neto, W.S., Costa, E.F., Marks, F.S.,
Rodenbusch, C.R., Canal, C.W. and Corbellini, L.G., (2013). Herd-level risk factors
for bovine viral diarrhea virus infection in dairy herds from Southern Brazil. Research
in veterinary science 95(3): 901-907.
Bahari, A., Gharekhani, J., Zandieh, M., Sadeghi-Nasab, A., Akbarein, H., Karimi-
Makhsous, A. and Yavari, M. (2013) Serological study of bovine herpes virus type 1
in dairy herds of Hamedan province, Iran. Veterinary Research Forum, 4 (2), 111-114.
Barber, K., Daugherty, H., Ander, S., Jefferson, V., Shack, L., Pechan, T., Nanduri, B.
and Meyer, F., (2017). Protein composition of the bovine herpesvirus 1.1 virion.
Journal of Veterinary sciences 4(1): 11.
Beer M., Konig P., Schielke G., Trapp S., (2003) Diagnostic markers in the prevention of
bovine herpesvirus type 1: possibilities and limitations, Berliner Und Munchener
Tierarztliche Wochenschrift 116:183-191.
Biswas Suman, Bandyopadhyay Samiran, Dimri Umesh & Patra H. Pabitra (2013)
Bovine herpesvirus-1 (BHV-1) – a re-emerging concern in livestock: a revisit to its
biology, epidemiology, diagnosis, and prophylaxis, Veterinary Quarterly 33 (2), 68-81,
Boelaert, F., Speybroeck, N., De Kruif, A., Aerts, M., Burzykowski, T., Molenberghs, G.
and Berkvens, D.L. (2005) Risk factors for bovine herpesvirus-1 seropositivity.
Preventive Veterinary Medicine 69 285-295.
45
Bosco Cowley, D. J., Clegg, T. A., Doherty, M. L. and More, S. J. (2011). Aspects of
bovine herpesvirus-1 infection in dairy and beef herds in Republic of Ireland. Acta
Veterinaria Scandinavica 53: 40.
Bowland S. L. and Shewen P. E. (2000) Bovine respiratory disease: commercial vaccines
currently available in Canada, The Canadian Veterinary Journal 41:33–48.
Callaby, R, Toye, PG, Jennings, A, Thumbi, SM, Coveter, JAW, Van Wyk, IC, Hanotte,
O, Mbole-Kariuki, MN, Bronsvoort, BMDC, Kruuk, LEB, Woolhouse, MEJ &
Kiara, H (2016) Seroprevalence of respiratory viral pathogens of indigenous calves in
Western Kenya' Research in Veterinary Science 108, 120-124.
Carbonero, A., Saa, L.R., Jara, D.V., García-Bocanegra, I., Arenas, A., Borge, C. and
Perea, A., (2011). Seroprevalence and risk factors associated to Bovine Herpesvirus 1
(BHV-1) infection in non-vaccinated dairy and dual purpose cattle herds in Ecuador.
Preventive veterinary medicine 100(1): 84-88.
Constable, P.D., Hinchcliff, K.W., Done, S.H., Grünberg, W., (2017). Infectious Bovine
Rhinotracheitis. Veterinary Medicine: A Textbook of the Diseases of Cattle, Horses,
Sheep, Pigs, and Goats, 11th edition. Elsevier, St. Louis, Missouri pp. 952–961.
Da Silva F. L., Gaudreault N. and Jones C. (2011). Cytoplasmic localized infected cell
protein 0 (bICP0) encoded by bovine herpesvirus 1 inhibits interferon promoter
activity and reduces IRF3 (interferon response factor 3) protein levels. Virus Research
160: 143–149.
Da Silva L. F., Sinani, D. and Jones C. (2012). ICP27 protein encoded by bovine
herpesvirus type 1 (bICP27) interferes with promoter activity of the bovine genes
encoding beta interferon 1 (IFN-1) and IFN-3. Virus Research 169: 162–168.
Dohoo, I. R., W. Martin, and H. Stryhn. (2009). Veterinary Epidemiologic Research. 2nd
ed. AVC Inc., Charlottetown, PEI, Canada.
46
Durham, P.J. and Hassard, L.E., (1990). Prevalence of antibodies to infectious bovine
rhinotracheitis, parainfluenza 3, bovine respiratory syncytial, and bovine viral diarrhea
viruses in cattle in Saskatchewan and Alberta. The Canadian Veterinary Journal
31(12): 815-820.
Fulton, R. W., Purdy, C. W., Confer, A.W., Saliki, J. T., Loan, R. W., Briggs, R. E.,
Burge, L. J., (2000) Bovine viral diarrhea viral infections in feeder calves with
respiratory disease: interactions with Pasteurella spp., parainfluenza-3 virus, and
bovine respiratory syncytial virus. Canadian Journal of Veterinary Research 64, 151–
159.Geraghty T., O'Neill R, Simon John More & O'Grady L. (2012) Dynamics of
individual animal Bovine Herpes Virus-1 antibody status on 9 commercial dairy herds.
Research in Veterinary Science 93 (1): 143 - 9.
Ghaemmaghami, Sh., Ahmadi, M., Deniko, A., Mokhberosafa, L. and Bakhshesh, M.
(2013) Serological study of BVDV and BHV-1 infections in industrial dairy herds of
Arak, Iran. Iranian Journal of Veterinary Science and Technology 5 (2), 53-61.
Ghirotti, M., Semproni, G., Meneghi, D., Mungaba, F.N., Nannini, D., Calzetta, G.,
Paganico, G., (1991) Sero-prevalences of selected cattle diseases in the Kafue flats of
Zambia. Veterinary Research Communication 15, 25–36.
González-Garcia, M.A., Arenas-Casas, A., Carbonero-Martínez, A., Borge-Rodríguez,
C., García-Bocanegra, I., Maldonado, J.L., Gómez, J.M. and Perea-Remujo, J.A.
(2009) Seroprevalence and risk factors associated with bovine herpesvirus type 1
(BHV-1) infection in non-vaccinated cattle herds in Andalusia (South of Spain). The
Spanish Journal of Agricultural Research 3, 550-554.
Gould, S., Cooper, V., Reichardt, N. and O’Connor, A. (2013). An evaluation of the
prevalence of bovine herpesvirus 1 abortions based on diagnostic submissions to five
47
U.S. based veterinary diagnostic laboratories. JThe Journal of Veterinary Diagnostic
Investigation 25: 243-247.
Graham D. A. (2013) Bovine herpes virus – 1 (BHV – 1) in cattle – a review with emphasis
on reproductive impacts and emergence of infection in Ireland and United Kingdom
Irish Veterinary Journal 66: 15.
Graham D., McShane J., Mawhinney K., McLaren I., Adair, B. and Merza M., (1998).
Evaluation of a single dilution ELISA system for detection of seroconversion to bovine
viral diarrhea virus, bovine respiratory syncytial virus, parainfluenza-3 virus, and
infectious bovine rhinotracheitis virus: comparison with testing by virus neutralization
and hemagglutination inhibition. Journal of Veterinary Diagnostic Investigation 10, 43–
48.
Griffin D., Chengappa, M. M., Kuszak J. and McVey D. S. (2010). Bacterial pathogens of
the bovine respiratory disease complex. Veterinary Clinics of North America: Food
Animal Practice 26, 381–39
Grissett, G. P., White, B. J., & Larson, R. L. (2015). Structured literature review of
responses of cattle to viral and bacterial pathogens causing bovine respiratory disease
complex. Journal of veterinary internal medicine 29 (3), 770-780.
Haji Hajikolaei, M. R. and Seyfi-Abad Shapouri, M. R. (2006). Seroepidemiological
Study of bovine herpes virus 1 (BHV-1) infection in cattle in Ahvaz. Iranian Veterinary
Journal 2: 2.
Handel, I.G., Willoughby, K., Land, F., Koterwas, B., Morgan, K.L., Tanya, V.N. and
Barend, M., (2011). Seroepidemiology of bovine viral diarrhoea virus (BVDV) in the
Adamawa region of Cameroon and use of the SPOT test to identify herds with PI
calves. PloS one, 6(7), p.e21620.
48
Henderson G., Zhang Y. and Jones C. (2005). The Bovine herpesvirus 1 gene encoding
infected cell protein 0 (bICP0) can inhibit interferon-dependent transcription in the
absence of other viral genes. Journal of General Virology 86: 2697–2702.
Jones, C. (2009) Regulation of Innate Immune Responses by Bovine Herpesvirus 1 and
Infected Cell Protein 0 (bICP0). Viruses 1, 255–275.
Kahrs R. F. (2001) Infectious bovine rhinotracheitis and infections pustular vulvovaginitis.
In: Viral Diseases of Cattle. 2nd ed. Ames, IA: Iowa State University Press 159–170.
Kenyanjui, M., Steiger, Y., Thorpe, W., (2007) Virus neutralizing Anitbodies to bovine
herpes virus type 1 (BHV-1), bovine viral diarrhoea (BVD) and rinderpest (RPV)
viruses in smallholder east African zebu cattle in coastal Kenya. Kenya Veterinarians
18, 21–24.
Keuser V, Schynts F, Detry B, Collard A, Robert B, Vanderplasschen A, Pastoret P,
Thirty E. (2004) Improved antigenic methods for differential of bovine diagnosis of
bovine, caprine, and corvine Alpha-herpesviruses related to bovine herpesvirus-1.
Journal of Clinical Microbiology 42:1228–1235.
King, A.M.Q., Adams, M.J., Carstens, E.B. and Lefkowitz, E.J. (2012) Virus Taxonomy:
Classification and Nomenclature of Viruses: Ninth Report of the International
Committee on Taxonomy of Viruses. San Diego, CA: Elsevier/Academic Press pp.
1010.
Koppers-Lalic, D., Rijsewijk, F. A. M., Verschuren, S. B. E., van Gaans-van den Brink,
J. A. M., Neisig, A., Ressing, M. E., Neefjes, J. and Wiertz, E. J. H. J. (2001). The
UL41-encoded virion host shutoff (vhs) protein and vhs- independent mechanisms are
responsible for down-regulation of MHC class I molecules by bovine herpesvirus 1.
Journal of General Virology 82, 2071–2081.
49
Kramps J.A., Banks M., Beer M., Kerkhofs P., Perrin M., Wellenberg G.J., van
Oirschot J.T., (2004) Evaluation of tests for antibodies against bovine herpesvirus 1
performed in national reference laboratories in Europe, Journal of Veterinary.
Microbiology 102:169–181.
Lehmann D., Sodoyer R., Leterme S., Crevat D., (2002) Improvement of serological
discrimination between herpesvirus-infected animals and animals vaccinated with
marker vaccines, Journal of Veterinary Microbiology 86:59–68.
Mahajan V, Banga H.S., Deva D., Filia G., Gupta A. (2013). Comparison of Diagnostic
Tests for Diagnosis of Infection Bovine Rhinotracheitis in Natural Cases of Bovine
Abortion. Journal of Comparative Pathology 149: 391-401.
Mahmoud MA, Mahmoud NA, Allam AM. (2009) Investigation on infectious bovine
rhinotracheitis in Egyptian cattle and buffaloes. Global Veterinaria 3: 335–340.
Mahony, T. J., (2010). Bovine herpesvirus: What is missing from our understanding of the
relationship between BoHV-1 and BoHV-5? The Veterinary Journal 2(184): 124-125.
Mainar-Jaime, R.C., Berzal-Herranz, B., Arias, P. and Rojo-Vázquez, F.A., (2001).
Epidemiological pattern and risk factors associated with bovine viral-diarrhoea virus
(BVDV) infection in a non-vaccinated dairy-cattle population from the Asturias region
of Spain. Preventive veterinary medicine 52(1): 63-73.
Majumder S., Ramakrishnan M. A. and Nandi S. (2015) Infectious Bovine
Rhinotracheitis: An Indian Perspective. International Journal of Current Microbiology
and Applied Sciences 4 (10): 844-858.
Makau D. N., VanLeeuwen J. A., Gitau G. K., Muraya J., McKenna S. L., Walton C.
and Wichtel J. J. (2018). Animal and management factors associated with weight gain
in dairy calves and heifers on smallholder dairy farms in Kenya. Preventive Veterinary
Medicine 161 60–68.
50
Mars M.H., de Jong M., Franken P., van Oirschot J.T., (2001) Efficacy of a live
glycoprotein E-negative bovine herpesvirus 1 vaccine in cattle in the field, Journal of
Vaccine 19:1924–1930.
Mars M.H., de Jong M.C., van Maanen C., Hage J.J., van Oirschot J.T., (2000) Airborne
transmission of bovine herpesvirus 1 infections in calves under field conditions.
Veterinary Microbiology 76: 1–13.
Mars, M. H., Bruschke, C. J., van Oirschot, J. T., (1999) Airborne transmission of BHV1,
BRSV, and BVDV among cattle is possible under experimental conditions. Veterinary
Microbiology 66 (3), 197 – 207.
Martin, S.W. and Bohac, J.G., (1986). The association between serological titers in
infectious bovine rhinotracheitis virus, bovine virus diarrhea virus, parainfluenza-3
virus, respiratory syncytial virus and treatment for respiratory disease in Ontario feedlot
calves. Canadian Journal of Veterinary Research, 50(3): 351.
McDermott, J.J., Kadohira, M., O'Callaghan, C.J. and Shoukri, M.M., (1997). A
comparison of different models for assessing variations in the sero-prevalence of
infectious bovine rhinotracheitis by farm, area and district in Kenya. Preventive
veterinary medicine 32(3-4): 219-234.
Moore S, Gunn M, and Walls D. (2000) Clinical and molecular analyses of bovine
herpesvirus 1 (BHV-1) isolates associated with disease in cattle in Ireland. Irish
Veterinary Journal 53: 89–93.
Muylkens, B., Thiry, J., Kirten, P., Schynts, F., Thiry, E., (2007) Bovine herpesvirus 1
infection and infectious bovine rhinotracheitis. Veterinary Records 38, 181–209.
Newcomer, B.W. and Givens, D. (2016) Diagnosis and control of viral diseases of
reproductive importance: Infectious Bovine Rhinotracheitis and Bovine Viral Diarrhea.
Veterinary Clinic North America Food Animal Practice, 34, 425-441.
51
O.I.E. (2008) Biological Standards Commission. OIE Manual of Diagnostic Tests and
Vaccines for Terrestrial Animals (mammals, birds and bees), 752-767.
O.I.E. (2010). Infectious bovine rhinotracheitis/infectious pustular vulvovaginitis. Chapter
2.4.13. In: Manual of Standards Diagnostic Tests and Vaccines 2010. Office
International des Epizooties.
Orjuela, J., Navarrete, M., Betancourt, A., Roqueme, L., Cortez, E. and Morrison, R.B.,
(1991). Salud y productividad en bovinos de la costa norte de Colombia. World Animal
Review, 69: 7-14.
Perrin B., Perrin M., Moussa A., Coudert M., (1996) Evaluation of a commercial gE
blocking ELISA test for detection of antibodies to infectious bovine rhinotracheitis
virus, Journal of Veterinary Records. 138:520.
Raaperi, K.; Orro, T.; Viltrop, A. (2014) Epidemiology and control of bovine herpesvirus 1
infection in Europe. Vet. J., 201, 249–256.
Radostits, O.M., Gay, C.C., Blood, D.C. and Hinchcliff, K.W. (2000) Infectious bovine
Rhinotracheitis (IBR, red nose), Bovine herpesvirus -1 (BHV-1) infection, 9th ed.,
W.B. Saunders, London. UK, 1173-1184.
Romero-Salas, D., Ahuja-Aguirre, C., Montiel-Palacios, F., García-Vázquez, Z., Cruz-
Romero, A. and Aguilar-Domínguez, M., (2013). Seroprevalence and risk factors
associated with infectious bovine rhinotracheitis in unvaccinated cattle in southern
Veracruz, Mexico. African journal of microbiology research, 7(17): 1716-1722.
Roshtkhari, F., Mohammadi, G. and Mayameei, A. (2012). Serological evaluation of
relationship between viral pathogens (BHV-1, BVDV, BRSV, PI-3V, and Adeno3) and
dairy calf pneumonia by indirect ELISA. Tropical Animal Health and Production 44(5):
1105-1110.
52
Saira K., Zhou Y. and Jones C. (2007). The infected cell protein 0 encoded by bovine
herpesvirus 1 (bICP0) induces degradation of interferon response factor 3 and,
consequently, inhibits beta interferon promoter activity. Journal of Virology 81: 3077–
3086.
Saira K., Zhou Y. and Jones C. (2009). The infected cell protein 0 encoded by bovine
herpesvirus 1 (bICP0) associates with interferon regulatory factor 7 and consequently
inhibits beta interferon promoter activity. Journal of Virology 83: 3977–3981.
Saravanajayam M., Kumanan K., Balasubramaniam A. (2015) Seroepidemiology of
infectious bovine rhinotracheitis infection in unvaccinated cattle, Journal of Veterinary
World, 8 (12): 1416-1419.
Segura-Correa J.C., Zapata-Campos C.C., Jasso-Obregón J.O., Martinez-Burnes J. and
López-Zavala R. (2016) Seroprevalence and risk factors associated with bovine
herpesvirus 1 and bovine viral diarrhea virus in North-Eastern Mexico, Open
Veterinary Journal, 6 (2): 143-149.
Seyfi Abad Shapouri, M., Ghiami Rad, M., Haji Hajikolaei, M. R., Mahmoodi, P.,
Karami, A., Daghari, M., (2016) Isolation of Bovine Herpesvirus-1 (BoHV-1) From
Latently Infected/Carrier Cattle in Ahvaz, Iranian Journal of Ruminants Health
Research, 1(1): 11-20.
Six, A., Banks, M., Engels, M., Bascuñana, C.R. and Ackermann, M., (2001). Latency
and reactivation of bovine herpesvirus 1 (BHV-1) in goats and of caprine herpesvirus 1
(CapHV-1) in calves. Archives of virology, 146(7): 1325-1335.
Snowder, G.D., Van Vleck, L.D., Cundiff, L.V. and Bennett, G.L., (2006). Bovine
respiratory disease in feedlot cattle: environmental, genetic, and economic factors.
Journal of animal science, 84(8): 1999-2008.
53
Solis-Calderon J.J., Segura-Correa V.M., Segura-Correa J.C., Alvarado-Islas A., (2003)
Seroprevalence of and risk factors for infectious bovine rhinotracheitis in beef cattle
herds of Yucatan, Mexico, Journal of Preventive Veterinary Medicine 57:199–208.
Straub OC. (1990) Infectious bovine rhinotracheitis virus. In: Dinter, Z. and Morein, B.,
editors. Virus infections of ruminants. Oxford: Elsevier Science Publishers BV; 71–
108.
Takiuchi, E., Medici, K., Alfieri, A., Alfieri, A., (2005) Bovine herpesvirus type 1 abortions
detected by a semi-nested PCR in Brazilian cattle herds. Research Veterinary Sciences
79, 85 88.
Van Oirschot J.T., Kaashoek M.J., MarisVeldhuis M.A., Weerdmeester K., Rijsewijk
F.A.M., (1997) An enzyme-linked immunosorbent assay to detect antibodies against
glycoprotein gE of bovine herpesvirus 1 allows differentiation between infected and
vaccinated cattle. Journal of Virology Methods 67:23–34.
Van Schaik G., (2001) Risk and economics of disease introduction to dairy farms, Tijdschr.
Diergeneeskd. 126:414–418 (in Dutch).
Van Schaik G., Schukken Y.H., Nielen M., Dijkhuizen A.A., Barkema H.W., Benedictus
G., (2002) Probability of and risk factors for introduction of infectious diseases into
Dutch SPF dairy farms: a cohort study. Journal of Preventive Veterinary Medicine.
54:279–289.
Viu, M.A., Dias, L.R.O., Lopes, D.T., Viu, A.F.M. and Ferraz, H.T. (2014) Infectious
bovine rhinotracheitis: Review. Pub. Vet. 8 (4).
Vonk Noordegraaf A., Labrovic A., Frankena K., Pfeiffer D.U., Nielen M., (2004)
Simulated hazards of losing infection-free status in a Dutch BHV1 model, Journal of
Preventive Veterinary Medicine 62:51–58.
54
Whetstone CA, and Evermann JF. (1988). Characterization of bovine herpesviruses
isolated from six sheep and four goats by restriction endonuclease analysis and
radioimmunoprecipitation. American Journal of Veterinary Research 49:781–785.
Winkler M.T., Doster A., Jones C., (1999) Bovine herpesvirus 1 can infect CD4 (+) T
lymphocytes and induce programmed cell death during acute infection of cattle, Journal
of Virology 73: 8657–8668.
Winkler, M.T., Doster, A. and Jones C. (2000) Persistence and reactivation of bovine
herpesvirus 1 in the tonsils of latently infected calves. Journal of Virology, 74, 5337-
5346.
Woodbine KA, Medley GF, Moore SJ, Ramirez-Villaescusa AM, Mason S, Green LE
(2009) A four-year longitudinal sero-epidemiological study of bovine herpesvirus type-
1 (BHV-1) in adult cattle in 107 unvaccinated herds in south west England. Veterinary
Research 5: 5.
Yan BF, Chao YJ, Chen Z, Tian KG, Wang CB, Lin XM, Chen HC, Guo AZ. (2008)
Serological survey of bovine herpesvirus type-1 infection in China. Veterinary
Microbiology 127: 136–141.
Yesilbag K, Bilge-dagalap S, Okur-gumusova S, Gunged B. (2003) Studies on herpesvirus
infections of goats in Turkey: Prevalence of antibodies to bovine herpesvirus 1. Revue
de médecine vétérinaire 154:772–774.
55
APPENDICES
Appendix 1: Descriptive statistics of continuous variables and proportion among the
principal farmers in 149 smallholder dairy farms, Naari area, Meru County, Kenya in
2018
Variable Category Frequency Proportion
Principal farmer Female 42 28.2
Male 84 56.4
Male and Female 23 15.4
Age of Males in the farm ≥ 45 years 73 49.0
< 45 years 76 51.0
Age of Females in the
farm
≥ 45 years 75 50.3
< 45 years 74 49.7
Marital status Married 125 83.9
single/Separated/Divorced/Widowed 24 16.1
Highest level of education
male in the farm
N/a/Primary school 30 20.2
High school 61 40.9
Tertiary school 58 38.9
Highest level of education
female in the farm
N/a/Primary school 24 16.1
High school 71 47.7
Tertiary school 54 36.2
Principal farmer with
Dairy training
No training 28 18.8
Trained on dairy 121 81.2
56
Appendix 2: Questionnaire for Management and Feeding Practices on Naari Smallholder
Dairy Farms
ASK QUESTIONS AS OPEN-ENDED (NOT GIVING ANSWERS); GIVE OPTIONS
IF NEEDED
Farmer Name: ________________________________________
Farm Number: _________________________________________
GPS lat: ________________________________________
GPS long: ________________________________________
Phone #: ________________________________________
Survey Visit Date: ________________________________________
Interviewer Initials: ________________________________________
1. Level of Education:
a) Primary [ ]
b) Secondary [ ]
c) College [ ]
d) University [ ]
e) N/A [ ]
2. Occupation:
a) Government employee
b) Self – employed
c)
3. Area of land owned: _______________ acres / hectares (circle units)
a) Percent of land used for crop and fodder production for cattle? [
] acres/ hectares
57
b) Area of land rented/used (unpaid): ________________ acres / hectares (circle
units)
c) Percent of land used for crop and fodder production for cattle?
d) Do you practice: Stall feeding ___________ or Semi – stall feed
_______________?
4. Have you attended any training on milk production in the last year? Y/N
If yes, what was this training about? ________________________________
5. What are the sources of information about dairy production in your farm?
a) Farmers training session [ ]
b) Internet [ ]
c) Other(specify)___________________________________________________
II. Feeding – Part A – Normal feeding:
6. Some feeds are only given seasonally. Over the last year, please check which of the
following you fed to your cattle (amounts not needed) and indicate the source
(purchased, shamba, neighbor, river, etc.).
Feed name Calves/Heifers Cows Source
a. Napier grass
b. Grass silage
c. Whole plant maize silage
c. Grass hay
d. Desmodium
e. Sweet potato vines
f. Other high protein forages – Lucerne, leucana, –
identify which one(s)
___________________________
g. Tree fodders – specify
_________________________
h. Maize stover
i. Banana leaves
j. Other fodder – specify (eg. Weeds) _________
k. Dairy meal
l. Wheat bran
m. Maize “Jam”
58
n. Vitamin/mineral powder
o. Vitamin/mineral block
p. Calf pellets/calf pencils. If yes, until what age?
q. Other feeds –specify (eg. Meal or cake)
r. Water available (always/sometimes ) A/S A/S
7. Do you usually feed dairy meal or grain to cows for the month before calving?
__YES __NO
If yes, do you increase the amount of dairy meal or grain during this month?
__YES __NO
8. Do you feed vitamins/minerals to cows during the month before calving?
___YES ___NO
If yes, what brand?
Brand: ____________ (from bag: Ca:P ratio: ___ Selenium amount & unit: ______)
If yes, how much is given to the cow? Amount (in spoons or grams per day):
9. For your cows, did you always have enough feeds over the last year? Yes_____
No_____
If no, which feeds were inadequate (check all that apply)?
Forages ___Grain or meals __Vitamin-minerals ___Water__ Other (specify)
10. Cattle deworming regime
a) How frequently do you normally deworm your cows?
Every ___ month’s ____when suspect it is a problem ___when not pregnant __
other?
b) How frequently do you normally deworm your calves/heifers?
Every ___ months ___ when suspect it is a problem ___ other (specify: ____)
59
c) Who normally deworms your cattle? Self: _ Vet service provider: ___ other:
___ who? __
d) What do you usually use to deworm your animals?
__________________________
11. Farm anima Deaths
a) How many calves died in the last year?
___________________________________
If some, from what causes __________________________________
b) How many heifers died in the last year?
__________________________________
If some, from what causes _______________________________________
c) How many cows died in the last year?
____________________________________
If some, from what causes
_____________________________________________
12. Veterinary Services
a) How far is the nearest vet service provider? ___________km
b) In the past year did a vet service provider visit your farm? Yes __ No __
If yes, for what reason? ___________________________________
c) Do you use an AI service? Y__N__
If yes, who provides your AI service? ___ Vet __Vet tech ___ AI tech ___
other_________________________
d) Do they come on time when you call them? Y [ ] N [ ]
e) Do they have the bulls you want to use? Y [ ] N [ ]
f) How do you decide what bull to use?
60
_____________________________________
13. Cow Stall Design and Management
a) How often do you remove manure from where the milking cows lie down?
a) Two times a day
b) Every day
c) Every other day
d) two times a week
e) Every week
f) Less than one time a week
b) How often do you add new bedding to where milking cows lie down?
a) Every other day
b) Every day
c) Two times a week
d) One time a week
e) Less than one time a week
c) How often do you add new bedding to where dry cows lie down?
a) Every other day
b) Every day
c) Two times a week
d) Once a week
e) Less than one time a week
d) How often do you trim your cow’s feet?
a) Every 4 months
b) Every 8 months
c) Every 12 months
61
d) Less often or never
14. For observation:
a) What kind of bedding is used where the milking cows lie down?
a) grass/hay b) straw c) sawdust d) crop waste e) soil f) none g) other (specify
_________)
b) Is the roof appropriate (observe – no holes, extends to cover udder area)? Yes
__No __
c) What is the type of the floor where the milking cows lie down?
1) Concrete 2) dirt 3) other (please specify: ________________)
d) Is the floor (observe – check all that apply):
1) Lumpy (have to lie on sticks, rocks, dirt chunks, etc.) Yes __ No
__
2) Hard (fails Knee impact test) Yes __ No
__
3) Wet in the udder area (fails the Knee wetness test) Yes __ No
__
e) Are there any sharp objects in the cow shed that may risk injuring the cattle (eg
nails)?
Yes __ No ___
f) Is water/urine/feces able to flow (by gravity) under udder where milking cows lie
down?
Yes ___No ____
15. Young-stock health and productivity checks (calves and heifers that have never
62
calved yet):
a) How do your calves usually receive their first colostrum? _____________________
b) Choose only ONE of the options that is MOST commonly used
Free choice suckle [ ]
Assisted suckle [ ]
Nursing bottle [ ]
Bucket [ ]
Other -specify: _______________________________________________________
c) How soon would most of your calves receive 4L of colostrum? Choose ONE answer
only
< 6 hours 6 - 12 hour’s _ 12 - 24 hours _ _ > 24 hours __ unknown ____
d) What do you usually do if a calf is weak and unable to drink colostrum during the first
day of life? Bottle feed ___Tube feed __ ___ Call Vet ______Other (specify) ______
e) Do you have a pen for pre-weaned calves? Y_____ N_____
If yes, type of flooring? ____________ 23c. Adequate roof? Y____ N____
Calf/Heifer #1
ID_______
Calf/Heifer#2
ID________
Calf/Heifer#3
ID_________
Calf/Heifer#4
ID_________
a. “Birthdate or Age (months)”
b. Sex
c. Breed
d. “First breeding date” (date or n/a)
d. “Latest breeding date” (date or n/a)
e.” Number of breeding’s to date” (# or
n/a)
f. “Had diarrhea” Y/N Y/N Y/N Y/N
g. “Had pneumonia” Y/N Y/N Y/N Y/N
h. “Had navel-ill” Y/N Y/N Y/N Y/N
63
i. Weight
j. Height
k. Body condition score
l. TPR/physical exam Normal / Abnormal?
(manure, feet, skin, lymph nodes, eyes,
rumen)
N / A N / A N / A N / A
m. Reproductive status (preg days
confirmed?)
Preg: Y/N Preg: Y/N Preg: Y/N Preg: Y/N
n. Month of last deworming
o. When last sprayed/dipped for ticks?
64
16. Health and Productivity of Cows
Examination of Cows: Cow1 (Q45)
ID_______
Cow2 (Q46)
ID______
Cow3 (Q47)
ID_______
Cow4 (Q48)
ID______
a. “Approximate age (years)”
b. Breed
c. “Number of calvings”
d. “Last calving date”
e. “First breeding date after last calving”
f. “Latest breeding date after last calving”
g. “Latest observed heat seen”
h. “Number of breedings for last
pregnancy”
i. “# of times used hormones for breeding”
j.” Current daily milk yield (kg/day)”
k. “Is this what she produced last week?” Y/N Y/N Y/N Y/N
m. “Abortion/stillbirths in last 12 months” Y/N Y/N Y/N Y/N
n. “Other disease (RP) in last 12
months”____
Y/N Y/N Y/N Y/N
o. Weight
p. Height
q. Body condition score
r. TPR/physical exam Normal / Abnormal?
(manure, feet, skin, lymph nodes, eyes,
rumen)
N / A N / A N / A N / A
s. Reproductive status (preg days
confirmed?)
Preg: Y/N Preg: Y/N Preg: Y/N Preg: Y/N
t. Normally get pregnancy confirmation? Y/N Y/N Y/N Y/N
u. Reproductive disease reported
v. Month of last deworming
w. When last sprayed/dipped for ticks?
x. Diseases vaccinated against (frequency)
65
a. In the last year, how frequently did your cows have an abrupt feed change? (For example, you
completely run out of one type of feed one day, such as Napier grass, so you switched to a
different type of feed the next day, such as weeds, banana leaves or maize stover) Choose ONE
Never ____Occasionally in the dry season ____ 1 time/month _____more than 1
time/month_____
b. In the last year, how frequently did your calves have an abrupt feed change? Choose ONE
Never ____Occasionally in the dry season ____ 1 time/month _____more than 1
time/month_____
c. What is the source of your cattle?
Buy as in-calf heifers [ ]
Buy as adult cows [ ]
Raise from young ones on my farm [ ]
d. Have you had a cow with difficult calving requiring assistance in the past? Yes No
If yes, who came to help? __________________________________________________
e. Have you had a cow that could not stand up before or after calving? Yes No
If yes, who came to help? ____________________________________________
f. Have you had a cow that had any other problem after calving? Yes No
If yes, what was it? _______________________________________________________
If yes, who came to help? __________________________________________________
g. Any other problems on the farm? Yes No
If yes, what was it? ________________________________________________________
If yes, did anyone come to help? _____________________________________________
If yes, still a problem? _____________________________________________________
17. Digital photo file range: ______________ to __________________